Pim kinase inhibitors and methods of their use

ABSTRACT

New compounds, compositions and methods of inhibition of kinase activity associated with tumorigenesis in a human or animal subject are provided. In certain embodiments, the compounds and compositions are effective to inhibit the activity of at least one serine/threonine kinase or receptor tyrosine kinase. The new compounds and compositions may be used either alone or in combination with at least one additional agent for the treatment of a serine/threonine kinase- or receptor tyrosine kinase-mediated disorder, such as cancer.

RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.12/449,701, filed Dec. 22, 2009, which is a national phase applicationunder 371 of International Application No. PCT/US08/055,724, filed Mar.3, 2008, which claims benefit to U.S. Provisional Application Nos.61/023,777, filed Jan. 25, 2008; and 60/892,444, filed Mar. 1, 2007. Allof the above listed applications are incorporated by reference herein intheir entirety.

FIELD OF THE INVENTION

The present invention relates to new compounds and their tautomers andstereoisomers, and pharmaceutically acceptable salts, esters,metabolites or prodrugs thereof, compositions of the new compoundstogether with pharmaceutically acceptable carriers, and uses of the newcompounds, either alone or in combination with at least one additionaltherapeutic agent, in the prophylaxis or treatment of cancer.

BACKGROUND

Infection with the Maloney retrovirus and genome integration in the hostcell genome results in development of lymphomas in mice. ProvirusIntegration of Maloney Kinase (PIM-Kinase) was identified as one of thefrequent proto-oncogenes capable of being transcriptionally activated bythis retrovirus integration event (Cuypers H T et al., “Murine leukemiavirus-induced T-cell lymphomagenesis: integration of proviruses in adistinct chromosomal region,” Cell 37(1):141-50 (1984); Selten G, etal., “Proviral activation of the putative oncogene Pim-1 in MuLV inducedT-cell lymphomas” EMBO J 4(7):1793-8 (1985)), thus establishing acorrelation between over-expression of this kinase and its oncogenicpotential. Sequence homology analysis demonstrated that there are 3highly homologous Pim-Kinases (Pim1, 2 & 3), Pim1 being theprotooncogene originally identified by retrovirus integration.Furthermore, transgenic mice over-expressing Pim1 or Pim2 show increasedincidence of T-cell lymphomas (Breuer M et al., “Very high frequency oflymphoma induction by a chemical carcinogen in pim-1 transgenic mice”Nature 340(6228):61-3 (1989)), while over-expression in conjunction withc-myc is associated with incidence of B-cell lymphomas (Verbeek S etal., “Mice bearing the E mu-myc and E mu-pim-1 transgenes developpre-B-cell leukemia prenatally” Mol Cell Biol 11(2):1176-9 (1991)).Thus, these animal models establish a strong correlation between Pimover-expression and oncogenesis in hematopoietic malignancies. Inaddition to these animal models, Pim over-expression has been reportedin many other human malignancies. Pim1, 2 & 3 over-expression isfrequently observed in many hematopoietic malignancies (Amson R et al.,“The human protooncogene product p33pim is expressed during fetalhematopoiesis and in diverse leukemias,” PNAS USA 86(22):8857-61 (1989);Cohen A M et al., “Increased expression of the hPim-2 gene in humanchronic lymphocytic leukemia and non-Hodgkin lymphoma,” Leuk Lymph45(S):951-5 (2004), Huttmann A et al., “Gene expression signaturesseparate B-cell chronic lymphocytic leukaemia prognostic subgroupsdefined by ZAP-70 and CD38 expression status,” Leukemia 20:1774-1782(2006)) and in prostate cancer (Dhanasekaran S M, et al., “Delineationof prognostic biomarkers in prostate cancer,” Nature 412(6849):822-6(2001); Cibull T L, et al., “Overexpression of Pim-1 during progressionof prostatic adenocarcinoma,” J Clin Pathol 59(3):285-8 (2006)), whileover-expression of Pim3 is frequently observed in hepatocellularcarcinoma (Fujii C, et al., “Aberrant expression of serine/threoninekinase Pim-3 in hepatocellular carcinoma development and its role in theproliferation of human hepatoma cell lines,” Int J Cancer 114:209-218(2005)) and pancreatic cancer (Li Y Y et al., “Pim-3, a proto-oncogenewith serine/threonine kinase activity, is aberrantly expressed in humanpancreatic cancer and phosphorylates bad to block bad-mediated apoptosisin human pancreatic cancer cell lines,” Cancer Res 66(13):6741-7(2006)).

Pim1, 2 & 3 are Serine/Threonine kinases normally function in survivaland proliferation of hematopoietic cells in response to growth factorsand cytokines. Cytokines signaling through the Jak/Stat pathway leads toactivation of transcription of the Pim genes and synthesis of theproteins. No further post-translational modifications are required forthe Kinase Pim activity. Thus, signaling down stream is primarilycontrolled at the transcriptional/translational and protein turnoverlevel. Substrates for Pim kinases include regulators of apoptosis suchas the Bcl-2 family member BAD (Aho T et al., “Pim-1 kinase promotesinactivation of the pro-apoptotic Bad protein by phosphorylating it onthe Ser112 gatekeeper site: FEBS Letters 571: 43-49 (2004)), cell cycleregulators such as p21^(WF1/CIP1) (Wang Z, et al., “Phosphorylation ofthe cell cycle inhibitor p21Cip1/WAF1 by Pim-1 kinase,” Biochim BiophysActa 1593:45-55 (2002)), CDC25A (1999), C-TAK (Bachmann M et al., “TheOncogenic Serine/Threonine Kinase Pim-1 Phosphorylates and Inhibits theActivity of Cdc25C-associated Kinase 1 (C-TAK1). A novel role for Pim-1at the G2/M cell cycle checkpoint,” J Biol Chem 179:48319-48328 (2004))and NuMA (Bhattacharya N, et al., “Pim-1 associates with proteincomplexes necessary for mitosis,” Chromosoma 111(2):80-95 (2002)) andthe protein synthesis regulator 4EBP1 (Hammerman P S et al., “Pim andAkt oncogenes are independent regulators of hematopoietic cell growthand survival,” Blood 105(11):4477-83 (2005)). The effects of Pim(s) inthese regulators are consistent with a role in protection from apoptosisand promotion of cell proliferation and growth. Thus, over-expression ofPim(s) in cancer is thought to play a role in promoting survival andproliferation of cancer cells and, therefore, their inhibitions shouldbe an effective way of treating cancers on which they areover-expressed. In fact several reports indicate that knocking downexpression of Pim(s) with siRNA results in inhibition of proliferationand cell death (Dai J M, et al., “Antisense oligodeoxynucleotidestargeting the serine/threonine kinase Pim-2 inhibited proliferation ofDU-145 cells,” Acta Pharmacol Sin 26(3):364-8 (2005); Fujii et al. 2005;Li et al. 2006). Furthermore, mutational activation of several well knowoncogenes in hematopoietic malignancies are thought exert its effects atleast in part through Pim(s). For example, targeted down regulation ofpim expression impairs survival of hematopoietic cells transformed byFlt3 and BCR/ABL (Adam et al. 2006). Thus, inhibitors to Pim1, 2 &3would be useful in the treatment of these malignancies. In addition to apotential role in cancer treatment and myeloproliferative diseases, suchinhibitor could be useful to control expansion of immune cells in otherpathologic condition such as autoimmune diseases, allergic reactions andin organ transplantation rejection syndromes. This notion is supportedby the findings that differentiation of Th1 Helper T-cells by IL-12 andIFN-α results in induction of expression of both Pim1 &2 (Aho T et al.,“Expression of human Pim family genes is selectively up-regulated bycytokines promoting T helper type 1, but not T helper type 2, celldifferentiation,” Immunology 116: 82-88 (2005)). Moreover, Pim(s)expression is inhibited in both cell types by the immunosuppressiveTGF-β (Aho et al. 2005). These results suggest that Pim kinases areinvolved in the early differentiation process of Helper T-cells, whichcoordinate the immunological responses in autoimmune diseases, allergicreaction and tissue transplant rejection.

In addition to PIM-Kinase, several other kinases have been shown to bedirectly involved in cancer, such as Flt3, KDR and PKCε. For example,several types of activating mutations in Flt3 are found in 20-30% ofpatients with Acute Myeloid Leukemia (AML). These activating mutationsare thought to be the most relevant transformation event on thesepatients and currently several Flt3 inhibitors are being tested for thetreatment on these patients in clinical trials (for a recent review seeTichenbrock L et al., “Emerging Flt3 kinase inhibitors in the treatmentof leukaemia,” Expert Opin Emerg Drugs 11:153-165 (2006)). KDR is one ofthe receptors for VEGF that plays a critical role in tumor angiogenesisand it's the target for the clinically validated bevacizumab drug (for arecent review see Ranieri G et al., “Vascular endothelial growth factor(VEGF) as a target of bevacizumab in cancer: from the biology to theclinic,” Curr Med Chem 13: 1845-1857 (2006)). Finally, over-expressionin NIH3T3 cells of PKCε has been shown to transform cell in vitro andpromote tumor formation in vivo (Perletti et al. Oncogene 12: 847(1996); Mischak et al., J Biol Chem 268: 6090 (1993)). In addition,over-expression of PKCε in the LNCaP cell line results in itstransformation to an androgen-independent tumor growth in nude nice (Wuet al., Cancer Research 62: 2423 (2002)). Furthermore, over-expressionof PKCε in transgenic mouse epithelium accelerates the development ofhighly malignant and rapidly metastasizing squamous cell carcinomas(Jansen et al., Cancer Research 61: 808 (2001)). Finally, clinically ifhas been observed that high PKCε expression in human tumors isassociated with poor disease-free and poor overall survival (Pan et al.,Cancer Research 65: 8366 (2005)). Thus, the compounds described herecould be useful in treating cancer by inhibiting these well validatedtargets of cancer drugs.

A continuing need exists for compounds that inhibit the proliferation ofcapillaries, inhibit the growth of tumors, treat cancer, modulate cellcycle arrest, and/or inhibit molecules such as Pim1, Pim2, Pim3, Flt3,KDR and PKCε, and pharmaceutical formulations and medicaments thatcontain such compounds. A need also exists for methods of administeringsuch compounds, pharmaceutical formulations, and medicaments to patientsor subjects in need thereof

SUMMARY

This summary is provided to introduce a selection of concepts in asimplified form that are further described below in the DetailedDescription. This summary is not intended to identify key features ofthe claimed subject matter, nor is it intended to be used as an aid indetermining the scope of the claimed subject matter.

New compounds, their tautomers and stereoisomers, and pharmaceuticallyacceptable salts thereof or esters having a solubility enhancingmoieties or prodrugs thereof are provided of the formula (I):

wherein,

X₁, X₂, X₃ and X₄ are independently selected from CR₂ and N; providedthat not more than two of X₁, X₂, X₃ and X₄ can be N;

Y is substituted or unsubstituted amino, alkoxy, cycloalkyl,heterocycloalkyl, aryl or heteroaryl;

Z₁, Z₂ and Z₃ are independently selected from CR₂ and N; provided thatnot more than one of Z₁, Z₂ and Z₃ can be N;

R₁ is selected from the group consisting of hydrogen, halo, alkyl,cycloalkyl, —CN, —NO₂, and —NHR₃;

each R₂ is independently selected from the group consisting of hydrogen,halo, hydroxyl, nitro, cyano, SO₃H and substituted or unsubstitutedalkyl, alkenyl, alkynyl, alkoxy, amino, aminocarbonyl,aminothiocarbonyl, aminocarbonylamino, aminothiocarbonylamino,aminocarbonyloxy, aminosulfonyl, aminosulfonyloxy, aminosulfonylamino,amidino, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxylester)oxy, sulfonyl, sulfonyloxy, thioacyl, thiol, alkylthio, aryl,heteroaryl, cycloalkyl, hetero cycloalkyl, partially saturatedcycloalkyl, aryloxy, heteroaryloxy, heterocyclyloxy, cycloalkyloxy,acyl, acylamino and acyloxy;

R₃ is selected from the group consisting of hydrogen, —CO—R₄ andsubstituted or unsubstituted alkyl, cycloalkyl, heterocyclyl, aryl andheteroaryl; and

R₄ is selected from the group consisting of alkyl, substituted alkyl,alkoxy, substituted alkoxy, amino, substituted amino, and alkylamino.

In other embodiments, new compounds are provided of the formula (II):

wherein,

Y is substituted or unsubstituted amino, cycloalkyl, heterocycloalkyl,aryl or heteroaryl;

Z₃ is selected from CR₂ and N;

R₁ is selected from the group consisting of hydrogen, halo, alkyl,cycloalkyl, —CN, —NO₂, and —NHR₃;

each R₂ is independently selected from the group consisting of hydrogen,halo, hydroxyl, nitro, cyano, SO₃H and substituted or unsubstitutedalkyl, alkenyl, alkynyl, alkoxy, amino, aminocarbonyl,aminothiocarbonyl, aminocarbonylamino, aminothiocarbonylamino,aminocarbonyloxy, aminosulfonyl, aminosulfonyloxy, aminosulfonylamino,amidino, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxylester)oxy, sulfonyl, sulfonyloxy, thioacyl, thiol, alkylthio, aryl,heteroaryl, cycloalkyl, hetero cycloalkyl, partially saturatedcycloalkyl, aryloxy, heteroaryloxy, heterocyclyloxy, cycloalkyloxy,acyl, acylamino and acyloxy;

R₃ is selected from the group consisting of hydrogen, —CO—R₄ andsubstituted or unsubstituted alkyl, cycloalkyl, heterocyclyl, aryl andheteroaryl; and

R₄ is selected from the group consisting of alkyl, substituted alkyl,alkoxy, substituted alkoxy, amino, substituted amino, and alkylamino

In other embodiments, new compounds are provided of the formula (III):

wherein,

Z₃ is selected from CR₂ and N;

R₁ is selected from the group consisting of hydrogen, halo, alkyl,cycloalkyl, —CN, —NO₂, and —NHR₃;

each R₂ is independently selected from the group consisting of hydrogen,halo, hydroxyl, nitro, cyano, SO₃H and substituted or unsubstitutedalkyl, alkenyl, alkynyl, alkoxy, amino, aminocarbonyl,aminothiocarbonyl, aminocarbonylamino, aminothiocarbonylamino,aminocarbonyloxy, aminosulfonyl, aminosulfonyloxy, aminosulfonylamino,amidino, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxylester)oxy, sulfonyl, sulfonyloxy, thioacyl, thiol, alkylthio, aryl,heteroaryl, cycloalkyl, hetero cycloalkyl, partially saturatedcycloalkyl, aryloxy, heteroaryloxy, heterocyclyloxy, cycloalkyloxy,acyl, acylamino and acyloxy;

R₃ is selected from the group consisting of hydrogen, —CO—R₄ andsubstituted or unsubstituted alkyl, cycloalkyl, heterocyclyl, aryl andheteroaryl; and

R₄ is selected from the group consisting of alkyl, substituted alkyl,alkoxy, substituted alkoxy, amino, substituted amino, and alkylamino

Also disclosed are compounds of the following formula (IV):

wherein,

R₁ is selected from the group consisting of hydrogen, halo, alkyl,cycloalkyl, —CN, —NO₂, and —NHR₃;

each R₂ is independently selected from the group consisting of hydrogen,halo, hydroxyl, nitro, cyano, SO₃H and substituted or unsubstitutedalkyl, alkenyl, alkynyl, alkoxy, amino, aminocarbonyl,aminothiocarbonyl, aminocarbonylamino, aminothiocarbonylamino,aminocarbonyloxy, aminosulfonyl, aminosulfonyloxy, aminosulfonylamino,amidino, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxylester)oxy, sulfonyl, sulfonyloxy, thioacyl, thiol, alkylthio, aryl,heteroaryl, cycloalkyl, hetero cycloalkyl, partially saturatedcycloalkyl, aryloxy, heteroaryloxy, heterocyclyloxy, cycloalkyloxy,acyl, acylamino and acyloxy;

R₃ is selected from the group consisting of hydrogen, —CO—R₄ andsubstituted or unsubstituted alkyl, cycloalkyl, heterocyclyl, aryl andheteroaryl; and

R₄ is selected from the group consisting of alkyl, substituted alkyl,alkoxy, substituted alkoxy, amino, substituted amino, and alkylamino

In other embodiments, new compounds are provided of formulas (I)-(IV),wherein Y is substituted or unsubstituted piperidinyl or piperazinyl. Inother embodiments, new compounds are provided of formulas (I)-(IV),wherein R₁ is hydrogen. In other embodiments, new compounds are providedof formulas (I)-(IV), wherein R₂ is independently selected from thegroup consisting of hydrogen, halo, hydroxyl, amino, nitro, cyano, SO₃Hand substituted or unsubstituted alkyl, aminoalkyl and phenyl.

In other aspects, the present invention provides methods for treatingPIM related disorders in a human or animal subject in need of suchtreatment comprising administering to said subject an amount of acompound of formula (I), (II), (III) or (IV) effective to PIM activityin the subject.

In other aspects, the present invention provides methods for treatingPIM related disorders in a human or animal subject in need of suchtreatment comprising administering to said subject an amount of acompound of formula (I), (II), (III) or (IV) effective to reduce orprevent tumor growth in the subject.

In yet other aspects, the present invention provides methods fortreating PIM related disorders in a human or animal subject in need ofsuch treatment comprising administering to said subject an amount of acompound of formula (I), (II), (III) or (IV) effective to reduce orprevent tumor growth in the subject in combination with at least oneadditional agent for the treatment of cancer.

In yet other aspects, the present invention provides therapeuticcompositions comprising at least one compound of formula (I), (II),(III) or (IV) in combination with one or more additional agents for thetreatment of cancer, as are commonly employed in cancer therapy.

The compounds of the invention are useful in the treatment of cancers,including hematopoietic malignancies, carcinomas (e.g., of the lungs,liver, pancreas, ovaries, thyroid, bladder or colon), melanoma, myeloiddisorders (e.g., myeloid leukemia, multiple myeloma anderythroleukemia), adenomas (e.g., villous colon adenoma), sarcomas(e.g., osteosarcoma), autoimmune diseases, allergic reactions and inorgan transplantation rejection syndromes.

In another aspect, the present invention relates to methods ofinhibiting at least one serine/threonine kinase in the Jak/Statsignaling pathway in a subject, or treating a biological conditionmediated by a serine/threonine kinase in the Jak/Stat signaling pathwayin a subject, comprising administering a therapeutic compositioncomprising at least one compound of formula (I), (II), (III) or (IV)effective to inhibit the Jak/Stat signaling pathway in the subject. Thetherapeutic compositions are useful for treating patients with a needfor such inhibitors (e.g., those suffering from cancer mediated byabnormal Jak/Stat signaling).

In another aspect, the present invention relates to methods ofinhibiting at least one serine/threonine kinase receptor selected fromthe group consisting of Pim1, Pim2 Pim3, Flt3, KDR and PKCε in asubject, or treating a biological condition mediated by at least one ofPim1, Pim2 Pim3, Flt3, KDR and PKCε comprising administering atherapeutic composition comprising at least one compound of formula (I),(II), (III) or (IV) effective to inhibit the kinase receptor in thesubject. The therapeutic compounds are useful for treating patients witha need for such inhibitors (e.g., those suffering from cancer mediatedby abnormal serine/threonine kinase receptor signaling).

The invention further provides compositions, methods of use, and methodsof manufacture as described in the detailed description of theinvention.

DETAILED DESCRIPTION

In accordance with one aspect of the present invention, compounds, theirtautomers and stereoisomers, and pharmaceutically acceptable saltsthereof or esters having a solubility enhancing moieties or prodrugsthereof are provided of the formula (I):

wherein,

X₁, X₂, X₃ and X₄ are independently selected from CR₂ and N; providedthat not more than two of X₁, X₂, X₃ and X₄ can be N;

Y is substituted or unsubstituted amino, alkoxy, cycloalkyl,heterocycloalkyl, aryl or heteroaryl;

Z₁, Z₂ and Z₃ are independently selected from CR₂ and N; provided thatnot more than one of Z₁, Z₂ and Z₃ can be N;

R₁ is selected from the group consisting of hydrogen, halo, alkyl,cycloalkyl, —CN, —NO₂, and —NHR₃;

each R₂ is independently selected from the group consisting of hydrogen,halo, hydroxyl, nitro, cyano, SO₃H and substituted or unsubstitutedalkyl, alkenyl, alkynyl, alkoxy, amino, aminocarbonyl,aminothiocarbonyl, aminocarbonylamino, aminothiocarbonylamino,aminocarbonyloxy, aminosulfonyl, aminosulfonyloxy, aminosulfonylamino,amidino, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxylester)oxy, sulfonyl, sulfonyloxy, thioacyl, thiol, alkylthio, aryl,heteroaryl, cycloalkyl, hetero cycloalkyl, partially saturatedcycloalkyl, aryloxy, heteroaryloxy, heterocyclyloxy, cycloalkyloxy,acyl, acylamino and acyloxy;

R₃ is selected from the group consisting of hydrogen, —CO—R₄ andsubstituted or unsubstituted alkyl, cycloalkyl, heterocyclyl, aryl andheteroaryl; and

R₄ is selected from the group consisting of alkyl, substituted alkyl,alkoxy, substituted alkoxy, amino, substituted amino, and alkylamino

In other embodiments, new compounds are provided of the formula (II):

wherein,

Y is substituted or unsubstituted amino, cycloalkyl, heterocycloalkyl,aryl or heteroaryl;

Z₃ is selected from CR₂ and N;

R₁ is selected from the group consisting of hydrogen, halo, alkyl,cycloalkyl, —CN, —NO₂, and —NHR₃;

each R₂ is independently selected from the group consisting of hydrogen,halo, hydroxyl, nitro, cyano, SO₃H and substituted or unsubstitutedalkyl, alkenyl, alkynyl, alkoxy, amino, aminocarbonyl,aminothiocarbonyl, aminocarbonylamino, aminothiocarbonylamino,aminocarbonyloxy, aminosulfonyl, aminosulfonyloxy, aminosulfonylamino,amidino, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxylester)oxy, sulfonyl, sulfonyloxy, thioacyl, thiol, alkylthio, aryl,heteroaryl, cycloalkyl, hetero cycloalkyl, partially saturatedcycloalkyl, aryloxy, heteroaryloxy, heterocyclyloxy, cycloalkyloxy,acyl, acylamino and acyloxy;

R₃ is selected from the group consisting of hydrogen, —CO—R₄ andsubstituted or unsubstituted alkyl, cycloalkyl, heterocyclyl, aryl andheteroaryl; and

R₄ is selected from the group consisting of alkyl, substituted alkyl,alkoxy, substituted alkoxy, amino, substituted amino, and alkylamino

In other embodiments, new compounds are provided of the formula (III):

wherein,

Z₃ is selected from CR₂ and N;

R₁ is selected from the group consisting of hydrogen, halo, alkyl,cycloalkyl, —CN, —NO₂, and —NHR₃;

each R₂ is independently selected from the group consisting of hydrogen,halo, hydroxyl, nitro, cyano, SO₃H and substituted or unsubstitutedalkyl, alkenyl, alkynyl, alkoxy, amino, aminocarbonyl,aminothiocarbonyl, aminocarbonylamino, aminothiocarbonylamino,aminocarbonyloxy, aminosulfonyl, aminosulfonyloxy, aminosulfonylamino,amidino, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxylester)oxy, sulfonyl, sulfonyloxy, thioacyl, thiol, alkylthio, aryl,heteroaryl, cycloalkyl, hetero cycloalkyl, partially saturatedcycloalkyl, aryloxy, heteroaryloxy, heterocyclyloxy, cycloalkyloxy,acyl, acylamino and acyloxy;

R₃ is selected from the group consisting of hydrogen, —CO—R₄ andsubstituted or unsubstituted alkyl, cycloalkyl, heterocyclyl, aryl andheteroaryl; and

R₄ is selected from the group consisting of alkyl, substituted alkyl,alkoxy, substituted alkoxy, amino, substituted amino, and alkylamino

Also disclosed are compounds of the following formula (IV):

wherein,

R₁ is selected from the group consisting of hydrogen, halo, alkyl,cycloalkyl, —CN, —NO₂, and —NHR₃;

each R₂ is independently selected from the group consisting of hydrogen,halo, hydroxyl, nitro, cyano, SO₃H and substituted or unsubstitutedalkyl, alkenyl, alkynyl, alkoxy, amino, aminocarbonyl,aminothiocarbonyl, aminocarbonylamino, aminothiocarbonylamino,aminocarbonyloxy, aminosulfonyl, aminosulfonyloxy, aminosulfonylamino,amidino, carboxyl, carboxyl ester, (carboxyl ester)amino, (carboxylester)oxy, sulfonyl, sulfonyloxy, thioacyl, thiol, alkylthio, aryl,heteroaryl, cycloalkyl, hetero cycloalkyl, partially saturatedcycloalkyl, aryloxy, heteroaryloxy, heterocyclyloxy, cycloalkyloxy,acyl, acylamino and acyloxy;

R₃ is selected from the group consisting of hydrogen, —CO—R₄ andsubstituted or unsubstituted alkyl, cycloalkyl, heterocyclyl, aryl andheteroaryl; and

R₄ is selected from the group consisting of alkyl, substituted alkyl,alkoxy, substituted alkoxy, amino, substituted amino, and alkylamino

In other embodiments, new compounds are provided of formulas (I)-(IV),wherein Y is substituted or unsubstituted piperidinyl or piperazinyl. Insome embodiments, new compounds are provided of formulas (I)-(IV),wherein X₁, X₃, and X₄ are —CH₂— and X₂ is —NH—. In some embodiments,new compounds are provided of formulas (I)-(IV), wherein X₅ is —CH₂—. Insome embodiments, new compounds are provided of formulas (I)-(IV),wherein X₆ is —CH(NH₂)—. In other embodiments, new compounds areprovided of formulas (I)-(IV), wherein R₁ is hydrogen. In otherembodiments, new compounds are provided of formulas (I)-(IV), wherein R₂is independently selected from the group consisting of hydrogen, halo,hydroxyl, amino, nitro, cyano, SO₃H and substituted or unsubstitutedalkyl, aminoalkyl and phenyl.

In other aspects, the present invention provides methods for treatingPIM related disorders in a human or animal subject in need of suchtreatment comprising administering to said subject an amount of acompound of formula (I), (II), (III) or (IV) effective to PIM activityin the subject.

In other aspects, the present invention provides methods for treatingPIM related disorders in a human or animal subject in need of suchtreatment comprising administering to said subject an amount of acompound of formula (I), (II), (III) or (IV) effective to reduce orprevent tumor growth in the subject.

In yet other aspects, the present invention provides methods fortreating PIM related disorders in a human or animal subject in need ofsuch treatment comprising administering to said subject an amount of acompound of formula (I), (II), (III) or (IV) effective to reduce orprevent tumor growth in the subject in combination with at least oneadditional agent for the treatment of cancer. A number of suitableanticancer agents to be used as combination therapeutics arecontemplated for use in the methods of the present invention. Indeed,the present invention contemplates, but is not limited to,administration of numerous anticancer agents such as: agents that induceapoptosis; polynucleotides (e.g., ribozymes); polypeptides (e.g.,enzymes); drugs; biological mimetics; alkaloids; alkylating agents;antitumor antibiotics; antimetabolites; hormones; platinum compounds;monoclonal antibodies conjugated with anticancer drugs, toxins, and/orradionuclides; biological response modifiers (e.g. interferons [e.g.IFN-α, etc.] and interleukins [e.g. IL-2, etc.], etc.); adoptiveimmunotherapy agents; hematopoietic growth factors; agents that inducetumor cell differentiation (e.g. all-trans-retinoic acid, etc.); genetherapy reagents; antisense therapy reagents and nucleotides; tumorvaccines; inhibitors of angiogenesis, and the like. Numerous otherexamples of chemotherapeutic compounds and anticancer therapies suitablefor coadministration with the disclosed compounds of formula (I), (II),(III) or (IV) are known to those skilled in the art.

In preferred embodiments, anticancer agents to be used in combinationwith compounds of the present invention comprise agents that induce orstimulate apoptosis. Agents that induce apoptosis include, but are notlimited to, radiation (e.g., W); kinase inhibitors (e.g., EpidermalGrowth Factor Receptor [EGFR] kinase inhibitor, Vascular Growth FactorReceptor [VGFR] kinase inhibitor, Fibroblast Growth Factor Receptor[FGFR] kinase inhibitor, Platelet-derived Growth Factor Receptor [PGFR]I kinase inhibitor, and Bcr-Abl kinase inhibitors such as STI-571,Gleevec, and Glivec]); antisense molecules; antibodies [e.g., Herceptinand Rituxan]; anti-estrogens [e.g., raloxifene and tamoxifen];anti-androgens [e.g., flutamide, bicalutamide, finasteride,aminoglutethamide, ketoconazole, and corticosteroids]; cyclooxygenase 2(COX-2) inhibitors [e.g., Celecoxib, meloxicam, NS-398, andnon-steroidal antiinflammatory drugs (NSAIDs)]; and cancerchemotherapeutic drugs [e.g., irinotecan (Camptosar), CPT-11,fludarabine (Fludara), dacarbazine (DTIC), dexamethasone, mitoxantrone,Mylotarg, VP-16, cisplatinum, 5-FU, Doxrubicin, Taxotere or taxol];cellular signaling molecules; ceramides and cytokines; and staurosprine,and the like.

In yet other aspects, the present invention provides therapeuticcompositions comprising at least one compound of formula (I), (II),(III) or (IV) in combination with a pharmaceutically acceptable carrier,and optionally with one or more additional agents for the treatment ofcancer, as are commonly employed in cancer therapy.

The compounds of the invention are useful in the treatment of cancers,including hematopoietic malignancies, carcinomas (e.g., of the lungs,liver, pancreas, ovaries, thyroid, bladder or colon), melanoma, myeloiddisorders (e.g., myeloid leukemia, multiple myeloma anderythroleukemia), adenomas (e.g., villous colon adenoma), sarcomas(e.g., osteosarcoma), autoimmune diseases, allergic reactions and inorgan transplantation rejection syndromes.

In another aspect, the present invention relates to methods ofinhibiting at least one serine/threonine kinase in the Jak/Statsignaling pathway in a subject, or treating a biological conditionmediated by a serine/threonine kinase in the Jak/Stat signaling pathwayin a subject, comprising administering a therapeutic compositioncomprising at least one compound of formula (I), (II), (III) or (IV)effective to inhibit the Jak/Stat signaling pathway in the subject. Thetherapeutic compositions are useful for treating patients with a needfor such inhibitors (e.g., those suffering from cancer mediated byabnormal Jak/Stat signaling).

In another aspect, the present invention relates to methods ofinhibiting at least one serine/threonine kinase receptor selected fromthe group consisting of Pim1, Pim2 Pim3, Flt3, KDR and PKCε in asubject, or treating a biological condition mediated by at least one ofPim1, Pim2 Pim3, Flt3, KDR and PKCε comprising administering atherapeutic composition comprising at least one compound of formula (I),(II), (III) or (IV) effective to inhibit the kinase receptor in thesubject. The therapeutic compounds are useful for treating patients witha need for such inhibitors (e.g., those suffering from cancer mediatedby abnormal serine/threonine kinase receptor signaling).

In another aspect, the present invention relates to methods ofinhibiting the activity of at least one kinase selected from the groupconsisting of Pim1, Pim2 Pim3, Flt3, KDR and PKCε in a subject, ortreating a biological condition mediated by at least one of Pim1, Pim2Pim3, Flt3, KDR and PKCε, in a human or animal subject in need of suchtreatment, comprising administering to the subject at least one compoundof formula (I), (II), (III) or (IV) in an amount effective to inhibitthe kinase in the subject. The therapeutic compounds are useful fortreating patients with a need for such inhibitors (e.g., those sufferingfrom cancer mediated by abnormal serine/threonine kinase receptorsignaling).

In other aspects, the present invention provides methods of manufactureof compounds of formula (I), (II), (III) or (IV) as described herein.

The invention further provides compositions, methods of use, and methodsof manufacture as described in the detailed description of theinvention.

“PIM inhibitor” is used herein to refer to a compound that exhibits anIC₅₀ with respect to PIM Kinase activity of no more than about 100 μMand more typically not more than about 50 μM, as measured in the PIMdepletion assays described hereinbelow.

The phrase “alkyl” refers to alkyl groups that do not containheteroatoms. Thus the phrase includes straight chain alkyl groups suchas methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl,decyl, undecyl, dodecyl and the like. The phrase also includes branchedchain isomers of straight chain alkyl groups, including but not limitedto, the following which are provided by way of example: —CH(CH₃)₂,—CH(CH₃)(CH₂CH₃), —CH(CH₂CH₃)₂, —C(CH₃)₃, —C(CH₂CH₃)₃, —CH₂CH(CH₃)₂,—CH₂CH(CH₃)(CH₂CH₃), —CH₂CH(CH₂CH₃)₂, —CH₂C(CH₃)₃, —CH₂C(CH₂CH₃)₃,—CH(CH₃)CH(CH₃)(CH₂CH₃), —CH₂CH₂CH(CH₃)₂, —CH₂CH₂CH(CH₃)(CH₂CH₃),—CH₂CH₂CH(CH₂CH₃)₂, —CH₂CH₂C(CH₃)₃, —CH₂CH₂C(CH₂CH₃)₃,—CH(CH₃)CH₂—CH(CH₃)₂, —CH(CH₃)CH(CH₃)CH(CH₃)₂,—CH(CH₂CH₃)CH(CH₃)CH(CH₃)(CH₂CH₃), and others. The phrase also includescyclic alkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, and cyclooctyl and such rings substituted withstraight and branched chain alkyl groups as defined above. Thus thephrase alkyl groups includes primary alkyl groups, secondary alkylgroups, and tertiary alkyl groups. Preferred alkyl groups includestraight and branched chain alkyl groups and cyclic alkyl groups having1 to 12 carbon atoms.

As used herein “loweralkyl” includes both substituted or unsubstitutedstraight or branched chain alkyl groups having from 1 to 6 carbon atoms.Representative loweralkyl groups include, for example, methyl, ethyl,propyl, isopropyl, n-butyl, tert-butyl, neopentyl, trifluoromethyl,pentafluoroethyl and the like. Loweralkyl groups may be substituted,such as with halo, hydroxy, amino, nitro and/or cyano groups, and thelike. Representative of halo-substituted and hydroxy-substitutedloweralkyl include chloromethyl, trichloromethyl, chloroethyl,hydroxyethyl, and the like. Other suitable substituted loweralkylmoieties include, for example, aralkyl, aminoalkyl, aminoaralkyl,carbonylaminoalkyl, alkylcarbonylaminoalkyl, arylcarbonylaminoalkyl,aralkylcarbonyl-aminoalkyl, aminoalkoxyalkyl and arylaminoalkyl.

“Loweralkoxy” as used herein refers to RO— wherein R is loweralkyl.Representative examples of loweralkoxy groups include methoxy, ethoxy,t-butoxy, trifluoromethoxy and the like.

As used herein, the term “halogen” or “halo” refers to chloro, bromo,fluoro and iodo groups. “Haloalkyl” refers to an alkyl radicalsubstituted with one or more halogen atoms. The term “haloloweralkyl”refers to a loweralkyl radical substituted with one or more halogenatoms. The term “haloalkoxy” refers to an alkoxy radical substitutedwith one or more halogen atoms. The term “haloloweralkoxy” refers to aloweralkoxy radical substituted with one or more halogen atoms.

“Amino” refers herein to the group —NH₂. The term “alkylamino” refersherein to the group —NRR′ where R and R′ are each independently selectedfrom hydrogen or a lower alkyl. The term “arylamino” refers herein tothe group —NRR′ where R is aryl and R′ is hydrogen, a lower alkyl, or anaryl. The term “aralkylamino” refers herein to the group —NRR′ where Ris a lower aralkyl and R′ is hydrogen, a loweralkyl, an aryl, or aloweraralkyl.

The term “alkoxyalkyl” refers to the group -alk₁-O-alk₂ where alk₁ isalkyl or alkenyl, and alk₂ is alkyl or alkenyl. The term“loweralkoxyalkyl” refers to an alkoxyalkyl where alk₁ is loweralkyl orloweralkenyl, and alk₂ is loweralkyl or loweralkenyl. The term“aryloxyalkyl” refers to the group -alkyl-O-aryl. The term“aralkoxyalkyl” refers to the group -alkylenyl-O-aralkyl, where aralkylis a loweraralkyl.

The term “aminocarbonyl” refers herein to the group —C(O)—NH₂.“Substituted aminocarbonyl” refers herein to the group —C(O)—NRR′ whereR is loweralkyl and R′ is hydrogen or a loweralkyl. In some embodiments,R and R′, together with the N atom attached to them may be takentogether to form a “heterocycloalkylcarbonyl” group. The term“arylaminocarbonyl” refers herein to the group —C(O)—NRR′ where R is anaryl and R′ is hydrogen, loweralkyl or aryl. “aralkylaminocarbonyl”refers herein to the group —C(O)—NRR′ where R is loweraralkyl and R′ ishydrogen, loweralkyl, aryl, or loweraralkyl.

“Aminosulfonyl” refers herein to the group —S(O)₂—NH₂. “Substitutedaminosulfonyl” refers herein to the group —S(O)₂—NRR′ where R isloweralkyl and R′ is hydrogen or a loweralkyl. The term“aralkylaminosulfonlyaryl” refers herein to the group-aryl-S(O)₂—NH-aralkyl, where the aralkyl is loweraralkyl.

“Carbonyl” refers to the divalent group —C(O)—. “Carboxy” refers to—C(═O)—OH. “Alkoxycarbonyl” refers to ester —C(═O)—OR wherein R isalkyl. “Loweralkoxycarbonyl” refers to ester —C(═O)—OR wherein R isloweralkyl. “Cycloalkyloxycarbonyl” refers to —C(═O)—OR wherein R iscycloalkyl. “Aryloxycarbonyl” refers to —C(═O)—OR wherein R is aryl.“Heterocyclyloxycarbonyl” refers to —C(═O)—OR wherein R is heterocyclyl.

The term “aralkoxycarbonyl” refers herein to the group —C(O)—O-aralkyl,where the aralkyl is loweraralkyl.

The term “sulfonyl” refers herein to the group —SO₂—. The term“sulfanyl” refers herein to the group —S—. “Loweralkylsulfonyl” refersto a substituted sulfonyl of the structure —SO₂R— in which R isloweralkyl. “Loweralkylsulfanyl” refers to a substituted sulfanyl of thestructure —SR— in which R is loweralkyl. Alkylsulfonyl and alkylsulfanylgroups employed in compounds of the present invention are typicallyloweralkylsulfonyl or loweralkylsulfanyl groups having from 1 to 6carbon atoms in its backbone structure. Thus, typical alkylsulfonyl andloweralkylsulfanyl groups employed in compounds of the present inventioninclude, for example, methylsulfonyl and methylsulfanyl (i.e., where Ris methyl), ethylsulfonyl and ethylsulfanyl (i.e., where R is ethyl),propylsulfonyl and propylsulfanyl (i.e., where R is propyl), and thelike. The term “arylsulfonyl” refers herein to the group —SO₂-aryl. Theterm “aralkylsulfonyl” refers herein to the group —SO₂-aralkyl, in whichthe aralkyl is loweraralkyl. The term “sulfonamido” refers herein to—SO₂NH₂.

As used herein, the term “carbonylamino” refers to the divalent group—NH—C(O)— in which the hydrogen atom of the amide nitrogen of thecarbonylamino group can be replaced a loweralkyl, aryl, or loweraralkylgroup. Such groups include moieties such as carbamate esters(—NH—C(O)—O—R) and amides —NH—C(O)—R, where R is a straight or branchedchain loweralkyl, cycloalkyl, or aryl or loweraralkyl.

“Cycloalkyl” refers to a mono- or polycyclic, heterocyclic orcarbocyclic alkyl substituent. Typical cycloalkyl substituents have from3 to 8 backbone (i.e., ring) atoms in which each backbone atom is eithercarbon or a heteroatom. The term “heterocycloalkyl” refers herein tocycloalkyl substituents that have from 1 to 5, and more typically from 1to 4 heteroatoms in the ring structure. Suitable heteroatoms employed incompounds of the present invention are nitrogen, oxygen, and sulfur.Representative heterocycloalkyl moieties include, for example,morpholino, piperazinyl, piperidinyl and the like. Carbocycloalkylgroups are cycloalkyl groups in which all ring atoms are carbon. Whenused in connection with cycloalkyl substituents, the term “polycyclic”refers herein to fused and non-fused alkyl cyclic structures.

The term “substituted heterocycle” or “heterocyclic group” orheterocycle as used herein refers to any 3- or 4-membered ringcontaining a heteroatom selected from nitrogen, oxygen, and sulfur or a5- or 6-membered ring containing from one to three heteroatoms selectedfrom the group consisting of nitrogen, oxygen, or sulfur; wherein the5-membered ring has 0-2 double bonds and the 6-membered ring has 0-3double bonds; wherein the nitrogen and sulfur atom may be optionallyoxidized; wherein the nitrogen and sulfur heteroatoms may be optionallyquarternized; and including any bicyclic group in which any of the aboveheterocyclic rings is fused to a benzene ring or another 5- or6-membered heterocyclic ring independently defined above. The term“heterocycle” thus includes rings in which nitrogen is the heteroatom aswell as partially and fully-saturated rings. Preferred heterocyclesinclude, for example: diazapinyl, pyrryl, pyrrolinyl, pyrrolidinyl,pyrazolyl, pyrazolinyl, pyrazolidinyl, imidazoyl, imidazolinyl,imidazolidinyl, pyridyl, piperidinyl, pyrazinyl, piperazinyl, N-methylpiperazinyl, azetidinyl, N-methylazetidinyl, pyrimidinyl, pyridazinyl,oxazolyl, oxazolidinyl, isoxazolyl, isoazolidinyl, morpholinyl,thiazolyl, thiazolidinyl, isothiazolyl, isothiazolidinyl, indolyl,quinolinyl, isoquinolinyl, benzimidazolyl, benzothiazolyl, benzoxazolyl,furyl, thienyl, triazolyl and benzothienyl.

Heterocyclic moieties can be unsubstituted or monosubstituted ordisubstituted with various substituents independently selected fromhydroxy, halo, oxo (C═O), alkylimino (RN═, wherein R is a loweralkyl orloweralkoxy group), amino, alkylamino, dialkylamino, acylaminoalkyl,alkoxy, thioalkoxy, polyalkoxy, loweralkyl, cycloalkyl or haloalkyl.

The heterocyclic groups may be attached at various positions as will beapparent to those having skill in the organic and medicinal chemistryarts in conjunction with the disclosure herein.

Where R is H or a heterocyclic substituent, as described herein.

Representative heterocyclics include, for example, imidazolyl, pyridyl,piperazinyl, azetidinyl, thiazolyl, furanyl, triazolyl benzimidazolyl,benzothiazolyl, benzoxazolyl, quinolinyl, isoquinolinyl, quinazolinyl,quinoxalinyl, phthalazinyl, indolyl, naphthpyridinyl, indazolyl, andquinolizinyl.

“Aryl” refers to optionally substituted monocyclic and polycyclicaromatic groups having from 3 to 14 backbone carbon or hetero atoms, andincludes both carbocyclic aryl groups and heterocyclic aryl groups.Carbocyclic aryl groups are aryl groups in which all ring atoms in thearomatic ring are carbon. The term “heteroaryl” refers herein to arylgroups having from 1 to 4 heteroatoms as ring atoms in an aromatic ringwith the remainder of the ring atoms being carbon atoms. When used inconnection with aryl substituents, the term “polycyclic aryl” refersherein to fused and non-fused cyclic structures in which at least onecyclic structure is aromatic, such as, for example, benzodioxozolo(which has a heterocyclic structure fused to a phenyl group, i.e.,

, naphthyl, and the like. Exemplary aryl moieties employed assubstituents in compounds of the present invention include phenyl,pyridyl, pyrimidinyl, thiazolyl, indolyl, imidazolyl, oxadiazolyl,tetrazolyl, pyrazinyl, triazolyl, thiophenyl, furanyl, quinolinyl,purinyl, naphthyl, benzothiazolyl, benzopyridyl, and benzimidazolyl, andthe like.

“Aralkyl” refers to an alkyl group substituted with an aryl group.Typically, aralkyl groups employed in compounds of the present inventionhave from 1 to 6 carbon atoms incorporated within the alkyl portion ofthe aralkyl group. Suitable aralkyl groups employed in compounds of thepresent invention include, for example, benzyl, picolyl, and the like.

Representative heteroaryl groups include, for example, those shownbelow. These heteroaryl groups can be further substituted and may beattached at various positions as will be apparent to those having skillin the organic and medicinal chemistry arts in conjunction with thedisclosure herein.

Representative heteroaryls include, for example, imidazolyl, pyridyl,piperazinyl, azetidinyl, thiazolyl, triazolyl benzimidazolyl,benzothiazolyl, and benzoxazolyl.

“Optionally substituted” or “substituted” refers to the replacement ofone or more hydrogen atoms with a monovalent or divalent radical.Suitable substitution groups include, for example, hydroxy, nitro,amino, imino, cyano, halo, thio, sulfonyl, thioamido, amidino, imidino,oxo, oxamidino, methoxamidino, imidino, guanidino, sulfonamido,carboxyl, formyl, loweralkyl, haloloweralkyl, loweralkylamino,haloloweralkylamino, loweralkoxy, haloloweralkoxy, loweralkoxyalkyl,alkylcarbonyl, aminocarbonyl, arylcarbonyl, aralkylcarbonyl,heteroarylcarbonyl, heteroaralkylcarbonyl, alkylthio, aminoalkyl,cyanoalkyl, aryl and the like.

The substitution group can itself be substituted. The group substitutedonto the substitution group can be carboxyl, halo; nitro, amino, cyano,hydroxy, loweralkyl, loweralkoxy, aminocarbonyl, —SR, thioamido, —SO₃H,—SO₂R or cycloalkyl, where R is typically hydrogen, hydroxyl orloweralkyl.

When the substituted substituent includes a straight chain group, thesubstitution can occur either within the chain (e.g., 2-hydroxypropyl,2-aminobutyl, and the like) or at the chain terminus (e.g.,2-hydroxyethyl, 3-cyanopropyl, and the like). Substituted substituentscan be straight chain, branched or cyclic arrangements of covalentlybonded carbon or heteroatoms.

It is understood that the above definitions are not intended to includeimpermissible substitution patterns (e.g., methyl substituted with fivefluoro groups or a halogen atom substituted with another halogen atom).Such impermissible substitution patterns are well known to the skilledartisan.

It will also be apparent to those skilled in the art that the compoundsof the invention, including the compounds of compounds of formulas (I),(II), (III) or (IV) or their stereoisomers, as well as thepharmaceutically acceptable salts, esters, metabolites and prodrugs ofany of them, may be subject to tautomerization and may therefore existin various tautomeric forms wherein a proton of one atom of a moleculeshifts to another atom and the chemical bonds between the atoms of themolecules are consequently rearranged. See, e.g., March, AdvancedOrganic Chemistry: Reactions, Mechanisms and Structures, Fourth Edition,John Wiley & Sons, pages 69-74 (1992). As used herein, the term“tautomer” refers to the compounds produced by the proton shift, and itshould be understood that the all tautomeric forms, insofar as they mayexist, are included within the invention.

The compounds of the invention, including the compounds of formulas (I),(II), (III) or (IV) or their tautomers, as well as the pharmaceuticallyacceptable salts, esters, metabolites and prodrugs of any of them, maycomprise asymmetrically substituted carbon atoms. Such asymmetricallysubstituted carbon atoms can result in the compounds of the inventionexisting in enantiomers, diastereomers, and other stereoisomeric formsthat may be defined, in terms of absolute stereochemistry, such as in(R)- or (S)-forms. As a result, all such possible isomers, individualstereoisomers in their optically pure forms, mixtures thereof, racemicmixtures (or “racemates”), mixtures of diastereomers, as well as singlediastereomers of the compounds of the invention are included in thepresent invention. The terms “S” and “R” configuration, as used herein,are as defined by the IUPAC 1974 RECOMMENDATIONS FOR SECTION E,FUNDAMENTAL STEREOCHEMISTRY, Pure Appi. Chem. 45:13-30 (1976). The termsα and β are employed for ring positions of cyclic compounds. The α-sideof the reference plane is that side on which the preferred substituentlies at the lower numbered position. Those substituents lying on theopposite side of the reference plane are assigned β descriptor. Itshould be noted that this usage differs from that for cyclicstereoparents, in which “a” means “below the plane” and denotes absoluteconfiguration. The terms α and β configuration, as used herein, are asdefined by the CHEMICAL ABSTRACTS INDEX GUIDE-APPENDIX IV (1987)paragraph 203.

As used herein, the term “pharmaceutically acceptable salts” refers tothe nontoxic acid or alkaline earth metal salts of the compounds ofFormulas (I), (II), (III) or (IV). These salts can be prepared in situduring the final isolation and purification of the compounds of Formulas(I), (II), (III) or (IV), or by separately reacting the base or acidfunctions with a suitable organic or inorganic acid or base,respectively. Representative salts include but are not limited to thefollowing: acetate, adipate, alginate, citrate, aspartate, benzoate,benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate,digluconate, cyclopentanepropionate, dodecylsulfate, ethanesulfonate,glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate,fumarate, hydrochloride, hydrobromide, hydroiodide,2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate,nicotinate, 2-naphthalenesulfonate, oxalate, pamoate, pectinate,persulfate, 3-phenylproionate, picrate, pivalate, propionate, succinate,sulfate, tartrate, thiocyanate, p-toluenesulfonate and undecanoate.Also, the basic nitrogen-containing groups can be quaternized with suchagents as loweralkyl halides, such as methyl, ethyl, propyl, and butylchloride, bromides, and iodides; dialkyl sulfates like dimethyl,diethyl, dibutyl, and diamyl sulfates, long chain halides such as decyl,lauryl, myristyl and stearyl chlorides, bromides and iodides, aralkylhalides like benzyl and phenethyl bromides, and others. Water oroil-soluble or dispersible products are thereby obtained.

Examples of acids which may be employed to form pharmaceuticallyacceptable acid addition salts include such inorganic acids ashydrochloric acid, sulfuric acid and phosphoric acid and such organicacids as oxalic acid, maleic acid, methanesulfonic acid, succinic acidand citric acid. Basic addition salts can be prepared in situ during thefinal isolation and purification of the compounds of formula (I), orseparately by reacting carboxylic acid moieties with a suitable basesuch as the hydroxide, carbonate or bicarbonate of a pharmaceuticallyacceptable metal cation or with ammonia, or an organic primary,secondary or tertiary amine Pharmaceutically acceptable salts include,but are not limited to, cations based on the alkali and alkaline earthmetals, such as sodium, lithium, potassium, calcium, magnesium, aluminumsalts and the like, as well as nontoxic ammonium, quaternary ammonium,and amine cations, including, but not limited to ammonium,tetramethylammonium, tetraethylammonium, methylamine, dimethylamine,trimethylamine, triethylamine, ethylamine, and the like. Otherrepresentative organic amines useful for the formation of base additionsalts include diethylamine, ethylenediamine, ethanolamine,diethanolamine, piperazine and the like.

As used herein, the term “pharmaceutically acceptable ester” refers toesters, which hydrolyze in vivo and include those that break downreadily in the human body to leave the parent compound or a saltthereof. Suitable ester groups include, for example, those derived frompharmaceutically acceptable aliphatic carboxylic acids, particularlyalkanoic, alkenoic, cycloalkanoic and alkanedioic acids, in which eachalkyl or alkenyl moiety advantageously has not more than 6 carbon atoms.Examples of particular esters include formates, acetates, propionates,butyrates, acrylates and ethylsuccinates.

The term “pharmaceutically acceptable prodrugs” as used herein refers tothose prodrugs of the compounds of the present invention which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of humans and lower animals without undue toxicity,irritation, allergic response, and the like, commensurate with areasonable benefit/risk ratio, and effective for their intended use, aswell as the zwitterionic forms, where possible, of the compounds of theinvention. The term “prodrug” refers to compounds that are rapidlytransformed in vivo to yield the parent compound of the above formula,for example by hydrolysis in blood. A thorough discussion is provided inT. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14of the A.C.S. Symposium Series, and in Edward B. Roche, ed.,Bioreversible Carriers in Drug Design, American PharmaceuticalAssociation and Pergamon Press, 1987, both of which are incorporatedherein by reference.

It will be apparent to those skilled in the art that the compounds ofthe invention, including the compounds of formulas (I), (II), (III) or(IV) or their tautomers, prodrugs and stereoisomers, as well as thepharmaceutically acceptable salts, esters and prodrugs of any of them,may be processed in vivo through metabolism in a human or animal body orcell to produce metabolites. The term “metabolite” as used herein refersto the formula of any derivative produced in a subject afteradministration of a parent compound. The derivatives may be producedfrom the parent compound by various biochemical transformations in thesubject such as, for example, oxidation, reduction, hydrolysis, orconjugation and include, for example, oxides and demethylatedderivatives. The metabolites of a compound of the invention may beidentified using routine techniques known in the art. See, e.g.,Bertolini, G. et al., J. Med. Chem. 40:2011-2016 (1997); Shan, D. etal., J. Pharm. Sci. 86(7):765-767; Bagshawe K., Drug Dev. Res.34:220-230 (1995); Bodor, N., Advances in Drug Res. 13:224-331 (1984);Bundgaard, H., Design of Prodrugs (Elsevier Press 1985); and Larsen, I.K., Design and Application of Prodrugs, Drug Design and Development(Krogsgaard-Larsen et al., eds., Harwood Academic Publishers, 1991). Itshould be understood that individual chemical compounds that aremetabolites of the compounds of formulas (I), (II), (III) or (IV) ortheir tautomers, prodrugs and stereoisomers, as well as thepharmaceutically acceptable salts, esters and prodrugs of any of them,are included within the invention.

The term “cancer” refers to cancer diseases that can be beneficiallytreated by the inhibition of Pim kinase, including, for example, solidcancers, such as carcinomas (e.g., of the lungs, pancreas, thyroid,ovarian, bladder, breast, prostate, or colon), melanomas, myeloiddisorders (e.g., myeloid leukemia, multiple myeloma anderythroleukemia), adenomas (e.g., villous colon adenoma) and sarcomas(e.g., osteosarcoma).

In other aspects, the present invention relates to the processes forpreparing the compounds of Formulas (I), (II) f(III) or IV, and to thesynthetic intermediates useful in such processes, as described in detailbelow.

Synthetic Methods

The compounds of the invention can be obtained through procedures knownto the skilled in the art. For example, as shown in Scheme 1,4-chloro,3-nitro pyridine can be reacted with a nucleophile yielding after nitroreduction a 4-substituted 3-amino pyridine I. Alternatively, 3-bromo4-nitro pyridine N-oxide can be reacted with a nucleophile, yieldingafter nitro and N-oxide reduction a 3-substituted 4-amino pyridine II.The substituted amino pyridines I and II can be acylated with carboxylicacids with the aid of coupling agents, or with acid halides or acidanhydrides yielding 3, 4 disubstituted pyridines III and IV. Compoundsof the invention containing 3, 4 disubstituted phenyls can be obtainedusing chemistry analogous to that in Scheme 1 when 3 halo 4 nitrobenzenes are the starting materials. Scheme 1.

Alternatively 3,4 disubstituted pyridines, as depicted in Scheme 2, canbe obtained by reacting halo nitro pyridines with boronic acids underSuzuki conditions, followed by nitro or nitro and N-oxide reductionyielding amino substituted pyridines V and VII. Subsequent amineacylation yields 3,4-disubstituted pyridines VI and VIII. Compounds ofthe invention containing 3,4-disubstituted phenyls can be obtained usingchemistry analogous to that in Scheme 2 when 3-halo, 4-nitro benzenesare the starting materials.

In an alternative manner, 3,4-disubstituted pyridines can be obtained asdepicted in Scheme 3. Formation of the bis-anion ofN-Boc-3-aminopyridine or N-pivaloyl-3-aminopyridine and reaction with anelectrophile yields 4-substituted, 3-N-protected amino pyridine IX.Acidic removal of the Boc or Piv protecting group and subsequentacylation yields the 3,4-substituted pyridine X. Compounds of theinvention containing 3,4-disubstituted phenyls can be obtained usingchemistry analogous to that in Scheme 3 when suitably protected anilinesare the starting materials.

Compounds of the invention containing 5-substituted, 4-aminoacylpyrimidones such as XII and XIII can be obtained as depicted in Scheme4. Nucleophilic substitution or Suzuki type coupling of 5-bromocytosinefollowed by N-acylation yields the 5-substituted, 4-aminoacylpyrimidones.

When the amide portion of substituted pyridines, benzenes or pyrimidonescontains a haloheteroaryl group, the substituted pyridines, benzenes orpyrimidones can be modified as depicted in Scheme 5. Direct carbonlinked groups (R′) can be attached to the heteroaryl group using Suzuki,Neghishi, Grignard or other organometallic methodologies. Alternativelynitrogen, oxygen, sulfur and carbon nucleophiles can be attached to theheteroaryl group utilizing standard methodologies including SnAr orBuchwald/Hartwig conditions.

The compounds of the invention are useful in vitro or in vivo ininhibiting the growth of cancer cells. The compounds may be used aloneor in compositions together with a pharmaceutically acceptable carrieror excipient. Suitable pharmaceutically acceptable carriers orexcipients include, for example, processing agents and drug deliverymodifiers and enhancers, such as, for example, calcium phosphate,magnesium stearate, talc, monosaccharides, disaccharides, starch,gelatin, cellulose, methyl cellulose, sodium carboxymethyl cellulose,dextrose, hydroxypropyl-β-cyclodextrin, polyvinylpyrrolidinone, lowmelting waxes, ion exchange resins, and the like, as well ascombinations of any two or more thereof. Other suitable pharmaceuticallyacceptable excipients are described in “Remington's PharmaceuticalSciences,” Mack Pub. Co., New Jersey (1991), incorporated herein byreference.

Effective amounts of the compounds of the invention generally includeany amount sufficient to detectably inhibit Pim activity by any of theassays described herein, by other Pim kinase activity assays known tothose having ordinary skill in the art or by detecting an inhibition oralleviation of symptoms of cancer.

The amount of active ingredient that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. It will beunderstood, however, that the specific dose level for any particularpatient will depend upon a variety of factors including the activity ofthe specific compound employed, the age, body weight, general health,sex, diet, time of administration, route of administration, rate ofexcretion, drug combination, and the severity of the particular diseaseundergoing therapy. The therapeutically effective amount for a givensituation can be readily determined by routine experimentation and iswithin the skill and judgment of the ordinary clinician.

For purposes of the present invention, a therapeutically effective dosewill generally be a total daily dose administered to a host in single ordivided doses may be in amounts, for example, of from 0.001 to 1000mg/kg body weight daily and more preferred from 1.0 to 30 mg/kg bodyweight daily. Dosage unit compositions may contain such amounts ofsubmultiples thereof to make up the daily dose.

The compounds of the present invention may be administered orally,parenterally, sublingually, by aerosolization or inhalation spray,rectally, or topically in dosage unit formulations containingconventional nontoxic pharmaceutically acceptable carriers, adjuvants,and vehicles as desired. Topical administration may also involve the useof transdermal administration such as transdermal patches orionophoresis devices. The term parenteral as used herein includessubcutaneous injections, intravenous, intramuscular, intrasternalinjection, or infusion techniques.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectable solutionor suspension in a nontoxic parenterally acceptable diluent or solvent,for example, as a solution in 1,3-propanediol. Among the acceptablevehicles and solvents that may be employed are water, Ringer's solution,and isotonic sodium chloride solution. In addition, sterile, fixed oilsare conventionally employed as a solvent or suspending medium. For thispurpose any bland fixed oil may be employed including synthetic mono- ordi-glycerides. In addition, fatty acids such as oleic acid find use inthe preparation of injectables.

Suppositories for rectal administration of the drug can be prepared bymixing the drug with a suitable nonirritating excipient such as cocoabutter and polyethylene glycols, which are solid at ordinarytemperatures but liquid at the rectal temperature and will thereforemelt in the rectum and release the drug.

Solid dosage forms for oral administration may include capsules,tablets, pills, powders, and granules. In such solid dosage forms, theactive compound may be admixed with at least one inert diluent such assucrose lactose or starch. Such dosage forms may also comprise, as isnormal practice, additional substances other than inert diluents, e.g.,lubricating agents such as magnesium stearate. In the case of capsules,tablets, and pills, the dosage forms may also comprise buffering agents.Tablets and pills can additionally be prepared with enteric coatings.

Liquid dosage forms for oral administration may include pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups, and elixirscontaining inert diluents commonly used in the art, such as water. Suchcompositions may also comprise adjuvants, such as wetting agents,emulsifying and suspending agents, cyclodextrins, and sweetening,flavoring, and perfuming agents.

The compounds of the present invention can also be administered in theform of liposomes. As is known in the art, liposomes are generallyderived from phospholipids or other lipid substances. Liposomes areformed by mono- or multi-lamellar hydrated liquid crystals that aredispersed in an aqueous medium. Any non-toxic, physiologicallyacceptable and metabolizable lipid capable of forming liposomes can beused. The present compositions in liposome form can contain, in additionto a compound of the present invention, stabilizers, preservatives,excipients, and the like. The preferred lipids are the phospholipids andphosphatidyl cholines (lecithins), both natural and synthetic. Methodsto form liposomes are known in the art. See, for example, Prescott, Ed.,Methods in Cell Biology, Volume XIV, Academic Press, New York, N.W., p.33 et seq. (1976).

While the compounds of the invention can be administered as the soleactive pharmaceutical agent, they can also be used in combination withone or more other agents used in the treatment of cancer. The compoundsof the present invention are also useful in combination with knowntherapeutic agents and anti-cancer agents, and combinations of thepresently disclosed compounds with other anti-cancer or chemotherapeuticagents are within the scope of the invention. Examples of such agentscan be found in Cancer Principles and Practice of Oncology, V. T. Devitaand S. Hellman (editors), 6th edition (Feb. 15, 2001), LippincottWilliams & Wilkins Publishers. A person of ordinary skill in the artwould be able to discern which combinations of agents would be usefulbased on the particular characteristics of the drugs and the cancerinvolved. Such anti-cancer agents include, but are not limited to, thefollowing: estrogen receptor modulators, androgen receptor modulators,retinoid receptor modulators, cytotoxic/cytostatic agents,antiproliferative agents, prenyl-protein transferase inhibitors, HMG-CoAreductase inhibitors and other angiogenesis inhibitors, inhibitors ofcell proliferation and survival signaling, apoptosis inducing agents andagents that interfere with cell cycle checkpoints. The compounds of theinvention are also useful when co-administered with radiation therapy.

Therefore, in one embodiment of the invention, the compounds of theinvention are also used in combination with known anticancer agentsincluding, for example, estrogen receptor modulators, androgen receptormodulators, retinoid receptor modulators, cytotoxic agents,antiproliferative agents, prenyl-protein transferase inhibitors, HMG-CoAreductase inhibitors, HIV protease inhibitors, reverse transcriptaseinhibitors, and other angiogenesis inhibitors.

Estrogen receptor modulators are compounds that interfere with orinhibit the binding of estrogen to the receptor, regardless ofmechanism. Examples of estrogen receptor modulators include, but are notlimited to, tamoxifen, raloxifene, idoxifene, LY353381, LY117081,toremifene, fulvestrant,4-[7-(2,2-dimethyl-1-oxopropoxy-4-methyl-2-[4-[2-(1-piperidinyl)ethoxy]phenyl]-2H-1-benzopyran-3-yl]-phenyl-2,2-dimethylpropanoate,4,4′-dihydroxybenzophenone-2,4-dinitrophenyl-hydra-zone, and SH646.

Androgen receptor modulators are compounds which interfere with orinhibit the binding of androgens to an androgen receptor. Representativeexamples of androgen receptor modulators include finasteride and other5α-reductase inhibitors, nilutamide, flutamide, bicalutamide, liarozole,and abiraterone acetate. Retinoid receptor modulators are compoundswhich interfere or inhibit the binding of retinoids to a retinoidreceptor. Examples of retinoid receptor modulators include bexarotene,tretinoin, 13-cis-retinoic acid, 9-cis-retinoic acid,α-difluoromethylornithine, LX23-7553, trans-N-(4′-hydroxyphenyl)retinamide, and N4-carboxyphenyl retinamide.

Cytotoxic and/or cytostatic agents are compounds which cause cell deathor inhibit cell proliferation primarily by interfering directly with thecell's functioning or inhibit or interfere with cell mytosis, includingalkylating agents, tumor necrosis factors, intercalators, hypoxiaactivatable compounds, microtubule inhibitors/microtubule-stabilizingagents, inhibitors of mitotic kinesins, inhibitors of kinases involvedin mitotic progression, antimetabolites; biological response modifiers;hormonal/anti-hormonal therapeutic agents, haematopoietic growthfactors, monoclonal antibody targeted therapeutic agents, topoisomeraseinhibitors, proteasome inhibitors and ubiquitin ligase inhibitors.Examples of cytotoxic agents include, but are not limited to, sertenef,cachectin, ifosfamide, tasonermin, lonidamine, carboplatin, altretamine,prednimustine, dibromodulcitol, ranimustine, fotemustine, nedaplatin,oxaliplatin, temozolomide, heptaplatin, estramustine, improsulfantosilate, trofosfamide, nimustine, dibrospidium chloride, pumitepa,lobaplatin, satraplatin, profiromycin, cisplatin, irofulven,dexifosfamide, cis-aminedichloro(2-methyl-pyridine)platinum,benzylguanine, glufos famide, GPX100, (trans, trans,trans)-bis-mu-(hexane-1,6-diamine)-mu-[diamine-platinum(II)]b is[diamine(chloro)platinum (II)]tetrachloride, diarizidinylspermine,arsenic trioxide,1-(11-dodecylamino-10-hydroxyundecyl)-3,7-dimethylxanthine, zorubicin,idarubicin, daunorubicin, bisantrene, mitoxantrone, pirarubicin,pinafide, valrubicin, amrubicin, antineoplaston,3′-deamino-3′-morpholino-13-deoxo-10-hydroxycamino-mycin, annamycin,galarubicin, elinafide, MEN10755, and4-demethoxy-3-deamino-3-aziridinyl-4-methylsulphonyl-daunorubicin (seeWO 00/50032). A representative example of a hypoxia activatable compoundis tirapazamine. Proteasome inhibitors include, but are not limited to,lactacystin and bortezomib. Examples of microtubuleinhibitors/microtubule-stabilizing agents include paclitaxel, vindesinesulfate, 3′,4′-didehydro-4′-deoxy-8′-norvincaleukoblastine, docetaxol,rhizoxin, dolastatin, mivobulin isethionate, auristatin, cemadotin,RPR109881, BMS184476, vinflunine, cryptophycin,2,3,4,5,6-pentafluoro-N-(3-fluoro-4-methoxyphenyl)benzene sulfonamide,anhydro-vinblastine,N,N-dimethyl-L-valyl-L-valyl-N-methyl-L-valyl-L-prolyl-L-proline-t-butyl-amide,TDX258, the epothilones (see for example U.S. Pat. Nos. 6,284,781 and6,288,237) and BMS188797. Representative examples of topoisomeraseinhibitors include topotecan, hycaptamine, irinotecan, rubitecan,6-ethoxypropionyl-3′,4′-O-exo-benzylidene-chartreusin,9-methoxy-N,N-dimethyl-5-nitropyrazolo[3,4,5-kl]acridine-2-(6H)propanamine,1-amino-9-ethyl-5-fluoro-2,3-dihydro-9-hydroxy-4-methyl-1H,12H-benzo[de]pyrano[3′,4′:b,7]-indolizino[1,2b]quinoline-10,13(9H,15H)dione, lurtotecan,7-[2-(N-isopropylamino)ethyl]-(20S)camptothecin, BNP 1350, BNPI1100,BN80915, BN80942, etoposide phosphate, teniposide, sobuzoxane,2′-dimethylamino-2′-deoxy-etoposide, GL331,N-[2-(dimethylamino)ethyl]-9-hydroxy-5,6-dimethyl-6H-pyrido[4,3-b]carbazole-1-carboxamide,asulacrine,(5a,5aB,8aa,9b)-9-[2-[N-[2-(dimethylamino)-ethyl]-N-methylamino]ethyl]-5-[4-hydroOxy-3,S-dimethoxyphenyl]-5,5a,6,8,8a,9-hexa-hydrofuro(3′,4′:6,7)naphtho(2,3-d)-1,3-dioxol-6-one,2,3-(methylenedioxy)-5-methyl-7-hydroxy-8-methoxybenzo[c]-phenanthridinium,6,9-bis[(2-aminoethyl)amino]benzo-[g]isoguinoline-5,10-dione,5-(3-aminopropylamino)-7,10-dihydroxy-2-(2-hydroxyethyl-aminomethyl)-6H-pyrazolo[4,5,1′-de]acridin-6-one,N-[1-[2(diethylamino)ethylamino]-7-methoxy-9-oxo-9H-thioxanthen-4-ylmethyl]formamide, N-(2-(dimethylamino)ethyl)-acridine-4-carboxamide,6-[[2-(dimethylamino)ethyl]amino]-3-hydroxy-7H-indeno[2,1-c]quinolin-7-one,and dimesna. Examples of inhibitors of mitotic kinesins, such as thehuman mitotic kinesin KSP, are described in PCT Publications WO 01/30768and WO 01/98278, WO 03/050,064 (Jun. 19, 2003), WO 03/050,122 (Jun. 19,2003), WO 03/049,527 (Jun. 19, 2003), WO 03/049,679 (Jun. 19, 2003), WO03/049,678 (Jun. 19, 2003) and WO 03/39460 (May 15, 2003) and pendingPCT Appl. Nos. US03/06403 (filed Mar. 4, 2003), US03/15861 (filed May19, 2003), US03/15810 (filed May 19, 2003),

US03/18482 (filed Jun. 12, 2003) and US03/18694 (filed Jun. 12, 2003).In an embodiment inhibitors of mitotic kinesins include, but are notlimited to inhibitors of KSP, inhibitors of MKLP1, inhibitors of CENP-E,inhibitors of MCAK, inhibitors of Kif14, inhibitors of Mphosphl andinhibitors of Rab6-KIFL.

Inhibitors of kinases involved in mitotic progression include, but arenot limited to, inhibitors of aurora kinase, inhibitors of Polo-likekinases (PLK) (e.g., inhibitors of PLK-1), inhibitors of bub-1 andinhibitors of bub-R1. Antiproliferative agents include antisense RNA andDNA oligonucleotides such as G3139, ODN698, RVASKRAS, GEM231, andINX3001, and antimetabolites such as enocitabine, carmofur, tegafur,pentostatin, doxifluridine, trimetrexate, fludarabine, capecitabine,galocitabine, cytarabine ocfosfate, fosteabine sodium hydrate,raltitrexed, paltitrexid, emitefur, tiazofurin, decitabine, nolatrexed,pemetrexed, nelzarabine, 2′-deoxy-2′-methylidenecytidine,2′-fluoromethylene-2′-deoxycytidine,N-[5-(2,3-dihydro-benzo-furyl)sulfonyl]-N′-(3,4-dichlorophenyl)urea,N6-[4-deoxy-4-[N2-[2(E),4(E)-tetradeca-dienoyl]glycylamino]-L-glycero-B-L-manno-heptopyranosyl]adenine,aplidine, ectein-ascidin, troxacitabine,4-[2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimidino[5,4-b]-[1,4]thiazin-6-yl-(S)-ethyl]-2,5-thienoyl-L-glutamicacid, aminopterin, 5-fluorouracil, alanosine,11-acetyl-8-(carbamoyloxymethyl)-4-formyl-6-methoxy-14-oxa-1,1-diaza-tetracyclo(7.4.1.0.0)-tetradeca-2,4,6-trien-9-ylacetic acid ester, swainsonine, lometrexol, dexrazoxane, methioninase,2′-cyano-2′-deoxy-N4-palmitoyl-1-B-D-arabino furanosyl cytosine and3-aminopyridine-2-carboxaldehyde thiosemicarbazone. Examples ofmonoclonal antibody targeted therapeutic agents include thosetherapeutic agents which have cytotoxic agents or radioisotopes attachedto a cancer cell specific or target cell specific monoclonal antibody.Examples include, for example, Bexxar. HMG-CoA reductase inhibitors areinhibitors of 3-hydroxy-3-methylglutaryl-CoA reductase. Compounds whichhave inhibitory activity for HMG-CoA reductase can be readily identifiedby using assays well-known in the art such as those described or citedin U.S. Pat. No. 4,231,938 and WO 84/02131. Examples of HMG-CoAreductase inhibitors that may be used include, but are not limited to,lovastatin (MEVACOR®, see U.S. Pat. Nos. 4,231,938, 4,294,926 and4,319,039), simvastatin (ZOCOR®, see U.S. Pat. Nos. 4,444,784, 4,820,850and 4,916,239), pravastatin (PRAVACHOL®; see U.S. Pat. Nos. 4,346,227,4,537,859, 4,410,629, 5,030,447 and 5,180,589), fluvastatin (LESCOL®;see U.S. Pat. Nos. 5,354,772, 4,911,165, 4,929,437, 5,189,164,5,118,853, 5,290,946 and 5,356,896) and atorvastatin (LIPITOR®; see U.S.Pat. Nos. 5,273,995, 4,681,893, 5,489,691 and 5,342,952). The structuralformulas of these and additional HMG-CoA reductase inhibitors that maybe used in the instant methods are described at page 87 of M. Yalpani,“Cholesterol Lowering Drugs”, Chemistry & Industry, pp. 85-89 (5 Feb.1996) and U.S. Pat. Nos. 4,782,084 and 4,885,314. In an embodiment, theHMG-CoA reductase inhibitor is selected from lovastatin and simvastatin.

Prenyl-protein transferase inhibitors are compounds which inhibit anyone or any combination of the prenyl-protein transferase enzymes,including farnesyl-protein transferase (FPTase), geranylgeranyl-proteintransferase type I (GGPTase-I), and geranylgeranyl-protein transferasetype-II (GGPTase-II, also called Rab GGPTase). Examples ofprenyl-protein transferase inhibiting compounds include(±)-6-[amino(4-chlorophenyl)(1-methyl-1H-imidazol-5-yl)methyl]-4-(3-chlorophenyl)-1-methyl-2(1H)-quinolinone,(−)-6-[amino(4-chlorophenyl)(1-methyl-1H-imidazol-5-yl)methyl]-4-(3-chlorophenyl)-1-methyl-2(1H)-quinolinone,(+)-6-[amino(4-chlorophenyl)(1-methyl-1H-imidazol-5-yl)methyl]-4-(3-chlorophenyl)-1-methyl-2(1H)-quinolinone,5(S)-n-butyl-1-(2,3-dimethylphenyl)-4-[1-(4-cyanobenzyl)-5-imidazolylmethyl-2-piperazinone,(S)-1-(3-chlorophenyl)-4-[1-(4-cyanobenzyl)-5-imidazolylmethyl]-5-[2-(ethanesulfonyl)methyl)-2-piperazinone,5(S)-n-butyl-1-(2-methylphenyl)-4-[1-(4-cyanobenzyl)-5-imidazolylmethyl]-2-piperazinone,1-(3-chlorophenyl)-4-[1-(4-cyanobenzyl)-2-methyl-5-imidazolylmethyl]-2-piperazinone,1-(2,2-diphenylethyl)-3-[N-(1-(4-cyanobenzyl)-1H-imidazol-5-ylethyl)carbamoyl]piperidine,4-{-[4-hydroxymethyl-4-(4-chloropyridin-2-ylmethyl)-piperidine-1-ylmethyl]-2-methylimidazol-1-ylmethyl}benzonitrile,4-{-5-[4-hydroxymethyl-4-(3-chlorobenzyl)-piperidine-1-ylmethyl]-2-methylimidazol-1-yl-methyl}benzonitrile,4-{3-[4-(2-oxo-2H-pyridin-1-yl)benzyl]-3H-imidazol-4-ylmethyl}-benzonitrile,4-{3-[4-(5-chloro-2-oxo-2H-[1,2]bipyridin-5′-ylmethyl]-3H-imidazol-4-yl-methyl}benzonitrile,4-{3-[4-(2-oxo-2H-[1,2′]bipyridin-5′-ylmethyl]-3H-imidazol4-yl-methyl}benzonitrile,4-{3-(2-oxo-1-phenyl-1,2-dihydropyridin-4-ylmethyl)-3H-imidazol-4-ylmethyl}benzonitrile,18,19-dihydro-19-oxo-5H,17H-6,1012,16-dimetheno-1H-imidazo[4,3-c][1,11,4]dioxaazacyclo-nonadecine-9-carbonitrile,(±)-19,20-dihydro-19-oxo-5H-18,21-ethano-12,14-etheno-6,10-metheno-22H-benzo[d]imidazo[4,3-k]-[1,6,9,12]oxatriaza-cyclooctadecine-9-carbonitrile,19,20-dihydro-19-oxo-5H,17H-18,21-ethano-6,10:12,16-dimetheno-22H-imidazo[3,4-h][1,8,11,14]oxatriazacycloeicosine-9-carbonitrile, and(.+−.)-19,20-dihydro-3-methyl-19-oxo-5H-18,21-ethano-12,14-etheno-6,10-metheno-22H-benzo[d]imidazo[4,3-k][1,6,9,12]oxa-triazacyclooctadecine-9-carbonitrile.Other examples of prenyl-protein transferase inhibitors can be found inthe following publications and patents: WO 96/30343, WO 97/18813, WO97/21701, WO 97/23478, WO 97/38665, WO 98/28980, WO 98/29119, WO95/32987, U.S. Pat. No. 5,420,245, U.S. Pat. No. 5,523,430, U.S. Pat.No. 5,532,359, U.S. Pat. No. 5,510,510, U.S. Pat. No. 5,589,485, U.S.Pat. No. 5,602,098, European Patent Publ. 0 618 221, European PatentPubl. 0 675 112, European Patent Publ. 0 604 181, European Patent Publ.0 696 593, WO 94/19357, WO 95/08542, WO 95/11917, WO 95/12612, WO95/12572, WO 95/10514, U.S. Pat. No. 5,661,152, WO 95/10515, WO95/10516, WO 95/24612, WO 95/34535, WO 95/25086, WO 96/05529, WO96/06138, WO 96/06193, WO 96/16443, WO 96/21701, WO 96/21456, WO96/22278, WO 96/24611, WO 96/24612, WO 96/05168, WO 96/05169, WO96/00736, U.S. Pat. No. 5,571,792, WO 96/17861, WO 96/33159, WO96/34850, WO 96/34851, WO 96/30017, WO 96/30018, WO 96/30362, WO96/30363, WO 96/31111, WO 96/31477, WO 96/31478, WO 96/31501, WO97/00252, WO 97/03047, WO 97/03050, WO 97/04785, WO 97/02920, WO97/17070, WO 97/23478, WO 97/26246, WO 97/30053, WO 97/44350, WO98/02436, and U.S. Pat. No. 5,532,359. For an example of the role of aprenyl-protein transferase inhibitor on angiogenesis see European J. ofCancer 35(9):1394-1401 (1999).

Angiogenesis inhibitors refers to compounds that inhibit the formationof new blood vessels, regardless of mechanism. Examples of angiogenesisinhibitors include, but are not limited to, tyrosine kinase inhibitors,such as inhibitors of the tyrosine kinase receptors Flt-1 (VEGFR1) andFlk-1/KDR (VEGFR2), inhibitors of epidermal-derived, fibroblast-derived,or platelet derived growth factors, MMP (matrix metalloprotease)inhibitors, integrin blockers, interferon-.alpha., interleukin-12,pentosan polysulfate, cyclooxygenase inhibitors, including nonsteroidalanti-inflammatories (NSAIDs) like aspirin and ibuprofen as well asselective cyclooxygenase-2 inhibitors like celecoxib and rofecoxib (PNAS89:7384 (1992); JNC1-69:475 (1982); Arch. Ophthalmol. 108:573 (1990);Anat. Rec., (238):68 (1994); FEBS Letters 372:83 (1995); Clin, Orthop.313:76 (1995); J. Mol. Endocrinol. 16:107 (1996); Jpn. J. Pharmacol.75:105 (1997); Cancer Res. 57:1625 (1997); Cell 93:705 (1998); Intl. J.Mol. Med. 2:715 (1998); J. Biol. Chem. 274:9116 (1999)), steroidalanti-inflammatories (such as corticosteroids, mineralocorticoids,dexamethasone, prednisone, prednisolone, methylpred, beta-methasone),carboxyamidotriazole, combretastatin A4, squalamine,6-O-chloroacetyl-carbonyl)-fumagillol, thalidomide, angiostatin,troponin-1, angiotensin II antagonists (see Fernandez et al., J. Lab.Clin. Med. 105:141-145 (1985)), and antibodies to VEGF (see, NatureBiotechnology, 17:963-968 (October 1999); Kim et al., Nature,362:841-844 (1993); WO 00/44777; and WO 00/61186). Other therapeuticagents that modulate or inhibit angiogenesis and may also be used incombination with the compounds of the instant invention include agentsthat modulate or inhibit the coagulation and fibrinolysis systems (seereview in Clin. Chem. La. Med. 38:679-692 (2000)). Examples of suchagents that modulate or inhibit the coagulation and fibrinolysispathways include, but are not limited to, heparin (see Thromb. Haemost.80:10-23 (1998)), low molecular weight heparins and carboxypeptidase Uinhibitors (also known as inhibitors of active thrombin activatablefibrinolysis inhibitor [TAFIa]) (see Thrombosis Res. 101:329-354(2001)). TAFIa inhibitors have been described in PCT Publication WO03/013,526 and U.S. Ser. No. 60/349,925 (filed Jan. 18, 2002). Theinvention also encompasses combinations of the compounds of theinvention with NSAIDs which are selective COX-2 inhibitors (generallydefined as those which possess a specificity for inhibiting COX-2 overCOX-1 of at least 100 fold as measured by the ratio of IC₅₀ for COX-2over IC₅₀ for COX-1 evaluated by cell or microsomal assays). Suchcompounds include, but are not limited to those disclosed in U.S. Pat.No. 5,474,995, issued Dec. 12, 1995, U.S. Pat. No. 5,861,419, issuedJan. 19, 1999, U.S. Pat. No. 6,001,843, issued Dec. 14, 1999, U.S. Pat.No. 6,020,343, issued Feb. 1, 2000, U.S. Pat. No. 5,409,944, issued Apr.25, 1995, U.S. Pat. No. 5,436,265, issued Jul. 25, 1995, U.S. Pat. No.5,536,752, issued Jul. 16, 1996, U.S. Pat. No. 5,550,142, issued Aug.27, 1996, U.S. Pat. No. 5,604,260, issued Feb. 18, 1997, U.S. Pat. No.5,698,584, issued Dec. 16, 1997, U.S. Pat. No. 5,710,140, issued Jan.20, 1998, WO 94/15932, published Jul. 21, 1994, U.S. Pat. No. 5,344,991,issued Jun. 6, 1994, U.S. Pat. No. 5,134,142, issued Jul. 28, 1992, U.S.Pat. No. 5,380,738, issued Jan. 10, 1995, U.S. Pat. No. 5,393,790,issued Feb. 20, 1995, U.S. Pat. No. 5,466,823, issued Nov. 14, 1995,U.S. Pat. No. 5,633,272, issued May 27, 1997, and U.S. Pat. No.5,932,598, issued Aug. 3, 1999, all of which are hereby incorporated byreference.

Representative inhibitors of COX-2 that are useful in the methods of thepresent invention include3-phenyl-4-(4-(methylsulfonyl)phenyl)-2-(5H)-furanone; and5-chloro-3-(4-methylsulfonyl)phenyl-2-(2-methyl-5-pyridinyl)pyridine.Compounds which are described as specific inhibitors of COX-2 and aretherefore useful in the present invention, and methods of synthesisthereof, can be found in the following patents, pending applications andpublications, which are herein incorporated by reference: WO 94/15932,published Jul. 21, 1994, U.S. Pat. No. 5,344,991, issued Jun. 6, 1994,U.S. Pat. No. 5,134,142, issued Jul. 28, 1992, U.S. Pat. No. 5,380,738,issued Jan. 10, 1995, U.S. Pat. No. 5,393,790, issued Feb. 20, 1995,U.S. Pat. No. 5,466,823, issued Nov. 14, 1995, U.S. Pat. No. 5,633,272,issued May 27, 1997, U.S. Pat. No. 5,932,598, issued Aug. 3, 1999, U.S.Pat. No. 5,474,995, issued Dec. 12, 1995, U.S. Pat. No. 5,861,419,issued Jan. 19, 1999, U.S. Pat. No. 6,001,843, issued Dec. 14, 1999,U.S. Pat. No. 6,020,343, issued Feb. 1, 2000, U.S. Pat. No. 5,409,944,issued Apr. 25, 1995, U.S. Pat. No. 5,436,265, issued Jul. 25, 1995,U.S. Pat. No. 5,536,752, issued Jul. 16, 1996, U.S. Pat. No. 5,550,142,issued Aug. 27, 1996, U.S. Pat. No. 5,604,260, issued Feb. 18, 1997,U.S. Pat. No. 5,698,584, issued Dec. 16, 1997, and U.S. Pat. No.5,710,140, issued Jan. 20, 1998. Other examples of angiogenesisinhibitors include, but are not limited to, endostatin, ukrain,ranpirnase, IM862,5-methoxy-4-[2-methyl-3-(3-methyl-2-butenyl)oxiranyl]-1-oxaspiro[2,5]oct-6-yl(chloroacetyl)carbamate,acetyldinanaline,5-amino-1-[[3,5-di-chloro-4-(4-chlorobenzoyl)phenyl]methyl]-1H-1,2,3-triazole-4-carboxamide,CM101, squalamine, combretastatin, RPI4610, NX₃₁₈₃₈, sulfatedmannopentanose phosphate,7,7-(carbonyl-bis[imino-N-methyl-4,2-pyrrolocarbonylimino[N-methyl-4,2-pyrrole]-carbonylimino]-bis-(1,3-naphthalenedisulfonate), and 3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone(SU5416).

Agents that interfere with cell cycle checkpoints are compounds thatinhibit protein kinases that transduce cell cycle checkpoint signals,thereby sensitizing the cancer cell to DNA damaging agents. Such agentsinclude inhibitors of ATR, ATM, the Chk1 and Chk2 kinases and cdk andcdc kinase inhibitors and are specifically exemplified by7-hydroxystaurosporin, flavopiridol, CYC202 (Cyclacel) and BMS-387032.

Inhibitors of cell proliferation and survival signaling pathway arepharmaceutical agents that inhibit cell surface receptors and signaltransduction cascades downstream of those surface receptors. Such agentsinclude inhibitors of inhibitors of EGFR (for example gefitinib anderlotinib), inhibitors of ERB-2 (for example trastuzumab), inhibitors ofIGFR, inhibitors of cytokine receptors, inhibitors of MET, inhibitors ofPI3K (for example LY294002), serine/threonine kinases (including but notlimited to inhibitors of Akt such as described in WO 02/083064, WO02/083139, WO 02/083140 and WO 02/083138), inhibitors of Raf kinase (forexample BAY-43-9006), inhibitors of MEK (for example CI-1040 andPD-098059) and inhibitors of mTOR (for example Wyeth CCI-779). Suchagents include small molecule inhibitor compounds and antibodyantagonists.

Apoptosis inducing agents include activators of TNF receptor familymembers (including the TRAIL receptors).

In certain presently preferred embodiments of the invention,representative agents useful in combination with the compounds of theinvention for the treatment of cancer include, for example, irinotecan,topotecan, gemcitabine, 5-fluorouracil, leucovorin carboplatin,cisplatin, taxanes, tezacitabine, cyclophosphamide, vinca alkaloids,imatinib (Gleevec), anthracyclines, rituximab, trastuzumab, as well asother cancer chemotherapeutic agents.

The above compounds to be employed in combination with the compounds ofthe invention will be used in therapeutic amounts as indicated in thePhysicians' Desk Reference (PDR) 47th Edition (1993), which isincorporated herein by reference, or such therapeutically useful amountsas would be known to one of ordinary skill in the art.

The compounds of the invention and the other anticancer agents can beadministered at the recommended maximum clinical dosage or at lowerdoses. Dosage levels of the active compounds in the compositions of theinvention may be varied so as to obtain a desired therapeutic responsedepending on the route of administration, severity of the disease andthe response of the patient. The combination can be administered asseparate compositions or as a single dosage form containing both agents.When administered as a combination, the therapeutic agents can beformulated as separate compositions, which are given at the same time ordifferent times, or the therapeutic agents, can be given as a singlecomposition.

Antiestrogens, such as tamoxifen, inhibit breast cancer growth throughinduction of cell cycle arrest, that requires the action of the cellcycle inhibitor p27Kip. Recently, it has been shown that activation ofthe Ras-Raf-MAP Kinase pathway alters the phosphorylation status ofp27Kip such that its inhibitory activity in arresting the cell cycle isattenuated, thereby contributing to antiestrogen resistance (Donovan etal., J. Biol. Chem. 276:40888, 2001). As reported by Donovan et al.,inhibition of MAPK signaling through treatment with MEK inhibitorchanged the phosphorylation status of p27 in hormone refactory breastcancer cell lines and in so doing restored hormone sensitivity.Accordingly, in one aspect, the compounds of formulas (I), (II), (III)and (IV) may be used in the treatment of hormone dependent cancers, suchas breast and prostate cancers, to reverse hormone resistance commonlyseen in these cancers with conventional anticancer agents.

In hematological cancers, such as chronic myelogenous leukemia (CML),chromosomal translocation is responsible for the constitutivelyactivated BCR-AB1 tyrosine kinase. The afflicted patients are responsiveto Gleevec, a small molecule tyrosine kinase inhibitor, as a result ofinhibition of Abl kinase activity. However, many patients with advancedstage disease respond to Gleevec initially, but then relapse later dueto resistance-conferring mutations in the Ab1 kinase domain. In vitrostudies have demonstrated that BCR-Av1 employs the Raf kinase pathway toelicit its effects. In addition, inhibiting more than one kinase in thesame pathway provides additional protection againstresistance-conferring mutations. Accordingly, in another aspect of theinvention, the compounds of formulas (I), (II), (III) and (IV) are usedin combination with at least one additional agent, such as Gleevec, inthe treatment of hematological cancers, such as chronic myelogenousleukemia (CML), to reverse or prevent resistance to the at least oneadditional agent.

In another aspect, the present invention relates to methods ofinhibiting at least one serine/threonine kinase in the Jak/Statsignaling pathway in a subject, or treating a biological conditionmediated by a serine/threonine kinase in the Jak/Stat signaling pathwayin a subject, comprising administering a therapeutic compositioncomprising at least one compound of formula (I), (II), (III) or (IV)effective to inhibit the activity of the at least one serine/threoninekinase in the Jak/Stat signaling pathway in the subject.

The therapeutic compositions in accordance with this aspect of theinvention are useful for treating patients with a need for suchinhibitors (e.g., those suffering from cancer mediated by abnormalJak/Stat signaling). Cancer types mediated by abnormal Jak/Statsignaling include, for example, melanoma, papillary cancer, thyroidcancer, ovarian cancer, colon cancer, pancreatic cancer, non-small celllung cancer (NSCLC), acute lymphoblastic leukemia (ALL), and acutemyeloid leukemia.

In one embodiment, the invention provides a method of inhibiting Pim1,Pim2 Pim3, Flt3, KDR or PKCε in a human or animal subject. The methodincludes administering an effective amount of a compound, or apharmaceutically acceptable salt thereof, of any of the embodiments ofcompounds of formula (I), (II), (III) or (IV) to a subject in needthereof.

The present invention will be understood more readily by reference tothe following examples, which are provided by way of illustration andare not intended to be limiting of the present invention.

Representative side chains for use in the compounds of the followingexamples may generally be prepared in accordance with the followingprocedures:

EXAMPLES

Referring to the examples that follow, compounds of the preferredembodiments were synthesized using the methods described herein, orother methods, which are known in the art.

The compounds and/or intermediates were characterized by highperformance liquid chromatography (HPLC) using a Waters Milleniumchromatography system with a 2695 Separation Module (Milford, Mass.).The analytical columns were reversed phase Phenomenex Luna C18-5μ,4.6×50 mm, from Alltech (Deerfield, Ill.). A gradient elution was used(flow 2.5 mL/min), typically starting with 5% acetonitrile/95% water andprogressing to 100% acetonitrile over a period of 10 minutes. Allsolvents contained 0.1% trifluoroacetic acid (TFA). Compounds weredetected by ultraviolet light (UV) absorption at either 220 or 254 nm.HPLC solvents were from Burdick and Jackson (Muskegan, Mich.), or FisherScientific (Pittsburgh, Pa.).

In some instances, purity was assessed by thin layer chromatography(TLC) using glass or plastic backed silica gel plates, such as, forexample, Baker-Flex Silica Gel 1B2-F flexible sheets. TLC results werereadily detected visually under ultraviolet light, or by employingwell-known iodine vapor and other various staining techniques.

Mass spectrometric analysis was performed on one of three LCMSinstruments: a Waters System (Alliance HT HPLC and a Micromass ZQ massspectrometer; Column. Eclipse XDB-C18, 2.1×50 mm; gradient: 5-95% (or35-95%, or 65-95% or 95-95%) acetonitrile in water with 0.05% TFA over a4 min period; flow rate 0.8 mL/min; molecular weight range 200-1500;cone Voltage 20 V; column temperature 40° C.), another Waters System(ACQUITY UPLC system and a ZQ 2000 system; Column. ACQUITY UPLC HSS—C18,1.8 um, 2.1×50 mm; gradient: 5-95% (or 35-95%, or 65-95% or 95-95%)acetonitrile in water with 0.05% TFA over a 1.3 min period; flow rate1.2 mL/min; molecular weight range 150-850; cone Voltage 20 V; columntemperature 50° C.) or a Hewlett Packard System (Series 1100 HPLC;Column. Eclipse XDB-C18, 2.1×50 mm; gradient: 5-95% acetonitrile inwater with 0.05% TFA over a 4 min period; flow rate 0.8 mL/min;molecular weight range 150-850; cone Voltage 50 V; column temperature30° C.). All masses were reported as those of the protonated parentions.

GCMS analysis was performed on a Hewlett Packard instrument (HP6890Series gas chromatograph with a Mass Selective Detector 5973; injectorvolume: 1 μL; initial column temperature: 50° C.; final columntemperature: 250° C.; ramp time: 20 minutes; gas flow rate: 1 mL/min;column: 5% phenyl methyl siloxane, Model No. HP 190915-443, dimensions:30.0 m×25 m×0.25 m).

Nuclear magnetic resonance (NMR) analysis was performed on some of thecompounds with a Varian 300 MHz NMR (Palo Alto, Calif.). The spectralreference was either TMS or the known chemical shift of the solvent.Some compound samples were run at elevated temperatures (e.g., 75° C.)to promote increased sample solubility.

The purity of some of the compounds is assessed by elemental analysis(Desert Analytics, Tucson, Ariz.).

Melting points are determined on a Laboratory Devices MeI-Temp apparatus(Holliston, Mass.).

Preparative separations are carried out using a Flash 40 chromatographysystem and KP-Sil, 60A (Biotage, Charlottesville, Va.), or by flashcolumn chromatography using silica gel (230-400 mesh) packing material,or by HPLC using a Waters 2767 Sample Manager, C-18 reversed phasecolumn, 30×50 mm, flow 75 mL/min. Typical solvents employed for theFlash 40 Biotage system and flash column chromatography aredichloromethane, methanol, ethyl acetate, hexane, acetone, aqueousammonia (or ammonium hydroxide), and triethyl amine Typical solventsemployed for the reverse phase HPLC are varying concentrations ofacetonitrile and water with 0.1% trifluoroacetic acid.

It should be understood that the organic compounds according to thepreferred embodiments may exhibit the phenomenon of tautomerism. As thechemical structures within this specification can only represent one ofthe possible tautomeric forms, it should be understood that thepreferred embodiments encompasses any tautomeric form of the drawnstructure.

It is understood that the invention is not limited to the embodimentsset forth herein for illustration, but embraces all such forms thereofas come within the scope of the above disclosure.

The examples below as well as throughout the application, the followingabbreviations have the following meanings. If not defined, the termshave their generally accepted meanings.

ABBREVIATIONS

DAST (diethylamino)sulfurtrifluoride DCM dichloromethane DIEAdiisopropylethylamine DMA dimethylacetamide DMAP 4-dimethylaminopyridineDME 1,2-dimethoxyethane DMF N,N-dimethylformamide DPPF1,1′-bis(diphenylphosphino)ferrocene EDC Ethyldimethylaminopropylazodicarboxylate hydrochloride EtOAc ethyl acetateEtOH Ethanol HOAT Hydroxyazabenzotriazole MeCN acetonitrile MeOHmethanol Na₂CO₃ sodium carbonate NaHCO₃ sodium bicarbonate NBSN-bromosuccinimide NMP N-methyl-2-pyrrolidone RT or rt room temperatureTDMSC1 tert-butyldimethylsilylchloride TEA triethylamine THFtetrahydrofuran

Example 1 Synthesis of 3-nitro-4-(piperidin-1-yl)pyridine

A solution of 4-chloro-3-nitropyridine (1.0 equiv.) and piperidine (2.0equiv.) in ethanol, at a concentration of 0.5 M, was stirred at rt for48 hours at which time the ethanol was removed in vacuo. The residue waspartitioned between EtOAc (300 mL) and Na₂CO_(3 (sat.)) (75 mL), waswashed further with H₂O (50 mL), NaCl_((sat)) (50 mL), was dried overMgSO₄, was filtered and the volatiles were removed in vacuo yielding3-nitro-4-(piperidin-1-yl)pyridine (95%). LCMS (m/z): 207.7 (MH⁺); LCR_(t)=1.60 min. ¹H NMR (CDCl₃): δ 8.80 (s, 1H), 8.31 (d, J=5.7, 1H),6.84 (d, J=6.3, 1H), 3.18-3.21 (m, 4H), 1.64-1.78 (m, 6H).

Example 2 Synthesis of tert-butyl1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate

The method of Example 1 was followed using 1 eq each of4-chloro-3-nitropyridine, 3-N-Boc-amino piperidine anddiisopropylethylamine yielding tert-butyl1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate, (89%). LCMS (m/z): 323.1(MH⁺); LC R_(t)=2.13 min.

Example 3 Synthesis of (R)-tert-butyl1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate

The method of Example 1 was followed using 1 eq each of4-chloro-3-nitropyridine, (R)-3-N-Boc-amino piperidine anddiisopropylethylamine yielding (R)-tert-butyl1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate, (99%). LCMS (m/z): 323.1(MH⁺); LC R_(t)=2.13 min.

Example 4 Synthesis of (S)-tert-butyl1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate

The method of Example 1 was followed using 1 eq each of4-chloro-3-nitropyridine, (S)-3-N-Boc-amino piperidine anddiisopropylethylamine yielding (S)-tert-butyl1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate, (99%). LCMS (m/z): 323.1(MH⁺); LC R_(t)=2.13 min.

Example 5 Synthesis of tert-butyl(1-(3-nitropyridin-4-yl)piperidin-3-yl)methylcarbamate

The method of Example 1 was followed using 1 eq each of4-chloro-3-nitropyridine, tert-butyl piperidin-3-ylmethylcarbamate anddiisopropylethylamine yielding tert-butyl(1-(3-nitropyridin-4-yl)piperidin-3-yl)methylcarbamate (99%). LCMS(m/z): 336.9 (MH⁺); LC R_(t)=2.27 min. ¹H NMR (CDCl₃): δ 8.80 (s, 1H),8.33 (d, J=6.0 Hz, 1H), 6.85 (d, J=6.3 Hz, 1H), 4.63 (bs, 1H), 3.28-3.46(m, 2H), 2.89-3.15 (m, 3H), 2.69-2.86 (m, 1H), 1.55-1.99 (m, 5H), 1.45(s, 9H).

Example 6 Synthesis of 1-(3-nitropyridin-4-yl)piperazine

The method of Example 1 was followed using 1 eq of4-chloro-3-nitropyridine and 10 eq of piperazine yielding tert-butyl1-(3-nitropyridin-4-yl)piperazine (99%). LCMS (m/z): 208.6 (MH⁺); LCR_(t)=0.42 min. ¹H NMR (CDCl₃): δ 8.83 (s, 1H), 8.37 (d, J=6.0, 1H),6.85 (d, J=6.0, 1H), 3.20-3.23 (m, 4H), 3.00-3.03 (m, 4H).

Example 7 Synthesis of tert-butyl1-(2-nitrophenyl)piperidin-3-ylcarbamate

The method of Example 1 was followed using 1-fluoro-2-nitrobenzene (1.0eq), 3-N-Boc-aminopiperidine (1.0 eq), and DIEA (2.0 eq) in EtOH at 50°C. for 48 hours yielding tert-butyl1-(2-nitrophenyl)piperidin-3-ylcarbamate (85%). LCMS (m/z): 322.2 (MH⁺);LC R_(t)=3.23 min.

Example 8 Synthesis of tert-butyl1-(2-nitrophenyl)piperidin-4-ylcarbamate

The method of Example 1 was followed using 1-fluoro-2-nitrobenzene (1.0eq), 4-N-Boc-aminopiperidine (1.2 eq), and TEA (2.0 eq) in EtOH at 55°C. for 48 hours yielding tert-butyl 1-(2-nitrophenyl)piperidin-4-ylcarbamate (100%). LCMS (m/z): 322.2 (MH⁺); LCR_(t)=3.15 min.

Example 9 Synthesis of tert-butyl4-(2-nitrophenyl)piperazine-1-carboxylate

The method of Example 1 was followed using 1-fluoro-2-nitrobenzene (1.0eq), 1-Boc-piperazine (1.2 eq), and TEA (2.0 eq) in EtOH at 55° C. for72 hours yielding tert-butyl 4-(2-nitro phenyl)piperazine-1-carboxylate(100%). LCMS (m/z): 308.1 (MH⁺); LC R_(t)=3.25 min.

Example 10 Synthesis of tert-butyl1-(3-nitropyridin-2-yl)piperidin-3-ylcarbamate

The method of Example 1 was followed using 2-chloro-3-nitropyridine (1.0eq), 3-N-Boc-aminopiperidine (1.2 eq), and DIEA (2.0 eq) yieldingtert-butyl 1-(3-nitropyridin-2-yl)piperidin-3-yl carbamate (95%). LCMS(m/z): 323.2 (MH⁺); LC R_(t)=3.00 min.

Example 11 Synthesis ofN,N-dimethyl-1-(3-nitropyridin-4-yl)piperidin-4-amine

The method of Example 1 was followed using 4-dimethylamino-piperidineyielding N,N-dimethyl-1-(3-nitropyridin-4-yl)piperidin-4-amine. LCMS(m/z): 251.2 (MH⁺).

Example 12 Synthesis of8-(3-nitropyridin-4-yl)-1,4-dioxa-8-azaspiro[4.5]decane

The method of Example 1 was followed using 1,4 dioxa-8-azaspiro[4.5]decane yielding 8-(3-nitropyridin-4-yl)-1,4-dioxa-8-azaspiro[4.5]decane. LCMS (m/z): 266.2 (MH⁺).

Example 13 Synthesis of tert-butyl4-(3-nitropyridin-4-yl)-1,4-diazepane-1-carboxylate

The method of Example 1 was followed using 1-Boc-homopiperazine yieldingtert-butyl 4-(3-nitropyridin-4-yl)-1,4-diazepane-1-carboxylate. LCMS(m/z): 293.3 (MH⁺);

Example 14 Synthesis ofN¹,N¹,N²-trimethyl-N²-(3-nitropyridin-4-yl)ethane-1,2-diamine

The method of Example 1 was followed using 4-chloro-3-nitropyridine (1.0eq), N¹,N¹,N²-trimethyl ethane-1,2-diamine (2.0 eq), and DIEA (2.0 eq)in EtOH yieldingN¹,N¹,N²-trimethyl-N²-(3-nitropyridin-4-yl)ethane-1,2-diamine which wasconcentrated and taken on as is. LCMS (m/z): 225.1 (MH⁺); LC R_(t)=0.574min.

Example 15 Synthesis of tert-butyl1-(3-nitropyridin-4-yl)pyrrolidin-3-ylcarbamate

The method of Example 1 was followed using 4-chloro-3-nitropyridine (1.0eq), tert-butyl pyrrolidin-3-ylcarbamate (2.0 eq), and DIEA (2.0 eq) inEtOH yielding tert-butyl 1-(3-nitropyridin-4-yl)pyrrolidin-3-ylcarbamate(95%) LCMS (m/z): 309.1 (MH⁺); LC R_(t)=1.922 min.

Example 16 Synthesis of (R)-tert-butyl[1-(3-nitropyridin-4-yl)pyrrolidin-2-yl]methylcarbamate

The method of Example 1 was followed using 4-chloro-3-nitropyridine (1.0eq), (R)-pyrrolidin-2-ylmethanamine (2.0 eq), and DIEA (2.0 eq) in EtOHyielding (R)-tert-butyl[1-(3-nitropyridin-4-yl)pyrrolidin-2-yl]methylcarbamate (95%) LCMS(m/z): 323.1 (MH⁺); LC R_(t)=1.855 min.

Example 17 Synthesis of 2-chloro-5-nitro-4-(piperidin-1-yl)pyrimidine

Method 1 was followed using 2,4-dichloro-5-nitropyrimidine (1.0 eq), andpiperidine (2.0 eq) in EtOH at 0° C. to rt, yielding after washing with1M citric acid and 1M HCl (to remove the bis addition product),2-chloro-5-nitro-4-(piperidin-1-yl)pyrimidine (67%) LCMS (m/z): 242.9(MH⁺); LC R_(t)=4.09 min.

Example 18 Synthesis of tert-butyl1-(3-fluoro-2-nitrophenyl)piperidin-4-ylcarbamate

Method 1 was followed using 1 eq each of 2,6-difluoronitrobenzene,4-(N-Boc-amino) piperidine, and TEA in EtOH yielding tert-butyl1-(3-fluoro-2-nitrophenyl)piperidin-4-ylcarbamate (93%). LCMS (m/z):340.1 (MH⁺); LC R_(t)=3.30 min.

Example 19 Synthesis of tert-butyl1-(5-fluoro-2-nitrophenyl)piperidin-4-ylcarbamate

Method 1 was followed using 1 eq each of 1,3-difluoro-4-nitrobenzene,4-(N-Boc-amino) piperidine, and TEA yielding tert-butyl1-(5-fluoro-2-nitrophenyl)-piperidin-4-ylcarbamate (93%). LCMS (m/z):340.1 (MH⁺); LC R_(t)=3.24 min.

Example 20 Synthesis of tert-butyl1-(4-fluoro-2-nitrophenyl)piperidin-4-ylcarbamate

Method 1 was followed using 2,5-difluoronitrobenzene (1.0 eq),4-(N-Boc-amino) piperidine (1.4 eq), and TEA (2.0 eq) at 55° C.overnight yielding tert-butyl1-(4-fluoro-2-nitrophenyl)piperidin-4-ylcarbamate (97%). LCMS (m/z):340.1 (MH⁺); LC R_(t)=3.28 min.

Example 21 Synthesis of tert-butyl1-(4-benzoyl-2-nitrophenyl)piperidin-3-ylcarbamate

Method 1 was followed using 4-chloro-3-nitrobenzophenone (1.0 eq),3-(N-Boc-amino) piperidine (1.1 eq), and TEA (2.0 eq) in NMP yieldingtert-butyl 1-(4-benzoyl-2-nitro phenyl)piperidin-3-ylcarbamate (90%).LCMS (m/z): 426.2 (MH⁺); LC R_(t)=3.49 min.

Example 22 Synthesis of tert-butyl1-(4-benzoyl-2-nitrophenyl)piperidin-4-ylcarbamate

Method 1 was followed using 4-chloro-3-nitrobenzophenone (1.0 eq),4-(N-Boc-amino) piperidine (1.1 eq), and TEA (2.0 eq) in NMP yieldingtert-butyl 1-(4-benzoyl-2-nitro phenyl)piperidin-4-ylcarbamate (95%).LCMS (m/z): 426.2 (MH⁺); LC R_(t)=3.46 min.

Example 23 Synthesis of tert-butyl4-(4-benzoyl-2-nitrophenyl)piperazine-1-carboxylate

Method 1 was followed using 4-chloro-3-nitrobenzophenone (1.0 eq),1-Boc-piperazine (1.1 eq), and TEA (2.0 eq) in NMP yielding tert-butyl4-(4-benzoyl-2-nitrophenyl)piperazine-1-carboxylate (93%). LCMS (m/z):412.2 (MH⁺); LC R_(t)=3.59 min.

Example 24 Synthesis of tert-butyl4-(4-acetyl-2-nitrophenyl)piperazine-1-carboxylate

Method 1 was followed using 4-chloro-3-nitroacetophenone (1.0 eq),t-Boc-piperazine (1.2 eq), and TEA (2.0 eq) at 55° C. overnight yieldingtert-butyl 4-(4-acetyl-2-nitro phenyl)piperazine-1-carboxylate (99%).LCMS (m/z): 350.1 (MH⁺); LC R_(t)=3.06 min.

Example 25 Synthesis of tert-butyl1-(4-acetyl-2-nitrophenyl)piperidin-4-ylcarbamate

Method 1 was followed using 4-chloro-3-nitroacetophenone (1.0 eq),4-(N-Boc-amino) piperidine (1.2 eq), and TEA (2.0 eq) at 55° C.overnight yielding tert-butyl1-(4-acetyl-2-nitrophenyl)piperidin-4-ylcarbamate (95%). LCMS (m/z):364.1 (MH⁺); LC R_(t)=2.99 min.

Example 26 Synthesis of tert-butyl1-(4-acetyl-2-nitrophenyl)piperidin-3-ylcarbamate

Method 1 was followed using 4-chloro-3-nitroacetophenone (1.0 eq),3-(N-Boc-amino) piperidine (1.2 eq), and TEA (2.0 eq) at 55° C.overnight tert-butyl 1-(4-acetyl-2-nitro phenyl)piperidin-3-ylcarbamate(99%). LCMS (m/z): 364.1 (MH⁺); LC R_(t)=3.03 min.

Example 27 Synthesis of tert-butyl1-(4-acetyl-2-nitrophenyl)piperidin-3-ylcarbamate

Method 1 was followed using 4-chloro-3-nitroanisole (1.0 eq),3-(N-Boc-amino) piperidine (1.2 eq), and TEA (2.0 eq) at 60° C. for 72hours yielding tert-butyl1-(4-methoxy-2-nitrophenyl)piperidin-3-ylcarbamate (50%). LCMS (m/z):352.1 (MH⁺); LC R_(t)=3.27 min.

Example 28 Synthesis of tert-butyl1-(4-methoxy-2-nitrophenyl)piperidin-4-ylcarbamate

Method 1 was followed using 4-chloro-3-nitroanisole (1.0 eq),4-(N-Boc-amino) piperidine (1.2 eq), and TEA (2.0 eq) at 60° C. for 72hours yielding tert-butyl1-(4-methoxy-2-nitrophenyl)piperidin-4-ylcarbamate (75%). LCMS (m/z):352.1 (MH⁺); LC R_(t)=3.22 min.

Example 29 Synthesis of tert-butyl4-(4-methoxy-2-nitrophenyl)piperazine-1-carboxylate

Method 1 was followed using 4-chloro-3-nitroanisole (1.0 eq),1-Boc-piperazine (1.2 eq), and TEA (2.0 eq) in NMP at 100° C. for 16hours yielding tert-butyl 4-(4-methoxy-2-nitrophenyl)piperazine-1-carboxylate (50%). LCMS (m/z): 338.2 (MH⁺); LCR_(t)=3.37 min.

Example 30 Synthesis of tert-butyl4-(4-chloro-2-nitrophenyl)piperazine-1-carboxylate

Method 1 was followed using 1 eq each of4-chloro-1-fluoro-2-nitrobenzene, 1-Boc-piperazine, and TEA yieldingtert-butyl 4-(4-chloro-2-nitrophenyl)piperazine-1-carboxylate (95%).LCMS (m/z): 342.0 (MH⁺); LC R_(t)=3.50 min.

Example 31 Synthesis of tert-butyl1-(4-chloro-2-nitrophenyl)piperidin-4-ylcarbamate

Method 1 was followed using 1 eq each of4-chloro-1-fluoro-2-nitrobenzene, 4-N-Boc-aminopiperidine, and TEAyielding tert-butyl 1-(4-chloro-2-nitrophenyl)piperidin-4-ylcarbamate(95%). LCMS (m/z): 356.1 (MH⁺); LC R_(t)=3.43 min.

Example 32 Synthesis of tert-butyl1-(4-chloro-2-nitrophenyl)piperidin-3-ylcarbamate

Method 1 was followed using 1 eq each of4-chloro-1-fluoro-2-nitrobenzene, 3-(N-Boc-amino) piperidine, and TEAyielding tert-butyl 1-(4-chloro-2-nitrophenyl)piperidin-4-ylcarbamate(97%). LCMS (m/z): 356.1 (MH⁺); LC R_(t)=3.47 min.

Example 33 Synthesis of tert-butyl4-(4-methyl-2-nitrophenyl)piperazine-1-carboxylate

Method 1 was followed using 4-fluoro-3-nitrotoluene (1.0 eq),1-Boc-piperazine (1.2 eq), and TEA (1.5 eq) at 55° C. for 48 hoursyielding tert-butyl 4-(4-methyl-2-nitrophenyl)piperazine-1-carboxylate(90%). LCMS (m/z): 322.1 (MH⁺); LC R_(t)=3.46 min.

Example 34 Synthesis of tert-butyl1-(4-methyl-2-nitrophenyl)piperidin-4-ylcarbamate

Method 1 was followed using 4-fluoro-3-nitrotoluene (1.0 eq),4-(N-Boc-amino) piperidine (1.2 eq), and TEA (1.5 eq) at 55° C. for 48hours yielding tert-butyl 1-(4-methyl-2-nitrophenyl)piperidin-4-ylcarbamate (87%). LCMS (m/z): 336.1 (MH⁺); LCR_(t)=3.32 min.

Example 35 Synthesis of tert-butyl1-(4-methyl-2-nitrophenyl)piperidin-3-ylcarbamate

Method 1 was followed using 4-fluoro-3-nitrotoluene (1.0 eq),3-(N-Boc-amino) piperidine (1.2 eq), and TEA (1.5 eq) at 55° C. for 48hours yielding tert-butyl1-(4-methyl-2-nitrophenyl)piperidin-3-ylcarbamate (87%). LCMS (m/z):336.1 (MH⁺); LC R_(t)=3.41 min.

Example 36 Synthesis of tert-butyl1-(2-nitro-4-(trifluoromethyl)phenyl)piperidin-3-ylcarbamate

Method 1 was followed using 1-fluoro-2-nitro-4-(trifluoromethyl)benzene(1.0 eq), 3-(N-Boc-amino) piperidine (1.2 eq), and TEA (1.5 eq) at 55°C. for 1 hour yielding tert-butyl1-(2-nitro-4-(trifluoromethyl)phenyl)piperidin-3-ylcarbamate (99%). LCMS(m/z): 390.1 (MH⁺); LC R_(t)=3.58 min.

Example 37 Synthesis of tert-butyl1-(2-nitro-4-(trifluoromethyl)phenyl)piperidin-4-ylcarbamate

Method 1 was followed using 1-fluoro-2-nitro-4-(trifluoromethyl)benzene(1.0 eq), 4-(N-Boc-amino) piperidine (1.2 eq), and TEA (2.0 eq) at 55°C. for 1 hour yielding tert-butyl1-(2-nitro-4-(trifluoromethyl)phenyl)piperidin-4-ylcarbamate (99%). LCMS(m/z): 390.1 (MH⁺); LC R_(t)=3.51 min.

Example 38 Synthesis of tert-butyl4-(2-nitro-4-(trifluoromethyl)phenyl)piperazine-1-carboxylate

Method 1 was followed using 1-fluoro-2-nitro-4-(trifluoromethyl)benzene(1.0 eq), 1-Boc-piperazine (1.2 eq), and TEA (2.0 eq) at 55° C. for 1hour yielding tert-butyl4-(2-nitro-4-(trifluoro-methyl)phenyl)piperazine-1-carboxylate (99%).LCMS (m/z): 376.1 (MH⁺); LC R_(t)=3.58 min.

Example 39 Synthesis of tert-butyl4-(5-methyl-2-nitrophenyl)piperazine-1-carboxylate

Method 1 was followed using 3-fluoro-4-nitrotoluene (1.0 eq),1-Boc-piperazine (1.2 eq), and TEA (2.0 eq) at 55° C. for 48 hoursyielding tert-butyl 4-(5-methyl-2-nitrophenyl)piperazine-1-carboxylate(97%). LCMS (m/z): 322.1 (MH⁺); LC R_(t)=3.43 min.

Example 40 Synthesis of tert-butyl1-(5-methyl-2-nitrophenyl)piperidin-4-ylcarbamate

Method 1 was followed using 3-fluoro-4-nitrotoluene (1.0 eq),4-(N-Boc-amino) piperidine (1.2 eq), and TEA (2.0 eq) at 55° C. for 48hours yielding tert-butyl1-(5-methyl-2-nitrophenyl)piperidin-4-ylcarbamate (97%). LCMS (m/z):336.1 (MH⁺); LC R_(t)=3.32 min

Example 41 Synthesis of tert-butyl1-(4-methyl-2-nitrophenyl)piperidin-3-ylcarbamate

Method 1 was followed using 3-fluoro-4-nitrotoluene (1.0 eq),3-(N-Boc-amino) piperidine (1.2 eq), and TEA (2.0 eq) at 55° C. for 48hours yielding tert-butyl1-(4-methyl-2-nitrophenyl)piperidin-3-ylcarbamate (98%). LCMS (m/z):336.1 (MH⁺); LC R_(t)=3.40 min.

Example 42 Synthesis of tert-butyl1-(4-cyano-2-nitrophenyl)piperidin-3-ylcarbamate

Method 1 was followed using 4-chloro-3-nitrobenzonitrile (1.0 eq),3-(N-Boc-amino) piperidine (1.0 eq), and DIEA (2.4 eq) at 55° C. for 24hours yielding tert-butyl1-(4-cyano-2-nitrophenyl)piperidin-3-ylcarbamate (95%). LCMS (m/z):347.2 (MH⁺); LC R_(t)=3.06 min.

Example 43 Synthesis of tert-butyl1-(2-nitro-4-(1H-pyrazol-5-yl)phenyl)piperidin-4-ylcarbamate

Method 1 was followed using 5-(4-chloro-3-nitrophenyl)-1H-pyrazole (1.0eq), 4-(N-Boc-amino) piperidine (1.1 eq), and TEA (2.0 eq) at 55° C. for24 hours yielding tert-butyl1-(2-nitro-4-(1H-pyrazol-5-yl)phenyl)piperidin-4-ylcarbamate. LCMS(m/z): 388.1 (MH⁺); LC R_(t)=2.84 min.

Example 44 Synthesis of tert-butyl1-(4-(methylsulfonyl)-2-nitrophenyl)piperidin-4-ylcarbamate

Method 1 was followed using 1-fluoro-4-(methylsulfonyl)-2-nitrobenzene(1.0 eq), 4-(N-Boc-amino) piperidine (1.1 eq), and TEA (2.0 eq) at 55°C. for 24 hours yielding tert-butyl1-(4-(methylsulfonyl)-2-nitrophenyl)piperidin-4-ylcarbamate. LCMS (m/z):400.1 (MH⁺); LC R_(t)=2.83 min.

Example 45 Synthesis of tert-butyl4-(4-(cyclopropanecarbonyl)-2-nitrophenyl)piperazine-1-carboxylate

Method 1 was followed using 4-chloro-3-nitrophenyl cyclopropyl ketone(1.0 eq), 1-Boc-piperazine (1.2 eq), and TEA (1.5 eq) at 55° C. for 48hours yielding tert-butyl4-(4-(cyclopropanecarbonyl)-2-nitrophenyl)piperazine-1-carboxylate

(98%). LCMS (m/z): 376.1 (MH⁺); LC R_(t)=3.33 min.

Example 46 Synthesis of tert-butyl1-(4-(cyclopropanecarbonyl)-2-nitrophenyl)piperidin-4-ylcarbamate

Method 1 was followed using 4-chloro-3-nitrophenyl cyclopropyl ketone(1.0 eq), 4-(N-Boc-amino)piperidine (1.2 eq), and TEA (1.5 eq) at 55° C.for 48 hours yielding tert-butyl1-(4-(cyclopropanecarbonyl)-2-nitrophenyl)piperidin-4-ylcarbamate (95%).LCMS (m/z): 390.1 (MH⁺); LC R_(t)=3.25 min.

Example 47 Synthesis of tert-butyl1-(4-(cyclopropanecarbonyl)-2-nitrophenyl)piperidin-3-ylcarbamate

Method 1 was followed using 4-chloro-3-nitrophenyl cyclopropyl ketone(1.0 eq), 3-(N-Boc-amino)piperidine (1.2 eq), and TEA (1.5 eq) at 55° C.for 48 hours yielding tert-butyl1-(4-(cyclopropanecarbonyl)-2-nitrophenyl)piperidin-3-ylcarbamate (96%).LCMS (m/z): 390.1 (MH⁺); LC R_(t)=3.28 min.

Example 48 Synthesis of tert-butyl2-(3-nitropyridin-4-yloxy)ethylcarbamate

To a cooled solution (0° C.) of tert-butyl 2-hydroxyethylcarbamate (1.1eq) in THF, NaH (1.3 eq) was added, stirred for 1 hr, and then4-chloro-3-nitropyridine (1.0 eq) was added. The reaction mixture wasstirred at RT overnight, poured into cold water, and extracted withEtOAc. Organic layer was dried over Na₂CO₃, filtered, and concentratedto yield tert-butyl 2-(3-nitropyridin-4-yloxy)ethylcarbamate. LCMS(m/z): 284.1 (MH⁺); LC R_(t)=2.09 min.

Method 2 Example 49 Synthesis of 4-(piperidin-1-yl)pyridin-3-amine

To a solution of 3-nitro-4-(piperidin-1-yl)pyridine (1.0 equiv.) inethanol, at a concentration of 0.1 M, was added 10% palladium on carbon(0.1 eq.). The resultant heterogeneous solution was put under anatmosphere of hydrogen and was stirred for 15 hours. At this time themixture was filtered through a pad of celite eluting with methanol. Thevolatiles were removed in vacuo yielding4-(piperidin-1-yl)pyridin-3-amine (93%) as an oil. LCMS (m/z): 178.0(MH⁺); LC R_(t)=1.68 min. ¹H NMR (CDCl₃): δ 8.01 (s, 1H), 7.96 (d,J=5.4, 1H), 6.78 (d, J=5.1, 1H), 3.64-3.74 (m, 2H), 2.86-2.94 (m, 4H),1.66-1.78 (m, 4H), 1.58-1.64 (m, 2H).

Example 50 Synthesis of tert-butyl1-(3-aminopyridin-4-yl)piperidin-3-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate was reduced yieldingtert-butyl 1-(3-aminopyridin-4-yl)piperidin-3-ylcarbamate, (65%). LCMS(m/z): 293.1 (MH⁺); LC R_(t)=2.10 min.

Example 51 Synthesis of (R)-tert-butyl1-(3-aminopyridin-4-yl)piperidin-3-ylcarbamate

Following the method of Example 49 (Method 2), (R)-tert-butyl1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate was reduced yielding(R)-tert-butyl 1-(3-aminopyridin-4-yl)piperidin-3-ylcarbamate, (89%).LCMS (m/z): 293.1 (MH⁺); LC R_(t)=2.08 min.

Example 52 Synthesis of (S)-tert-butyl1-(3-aminopyridin-4-yl)piperidin-3-ylcarbamate

Following the method of Example 49 (Method 2), (S)-tert-butyl1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate was reduced yielding(S)-tert-butyl 1-(3-aminopyridin-4-yl)piperidin-3-ylcarbamate, (78%).LCMS (m/z): 293.1 (MH⁺); LC R_(t)=2.08 min.

Example 53 Synthesis of tert-butyl(1-(3-aminopyridin-4-yl)piperidin-3-yl)methylcarbamate

Following the method of Example 49 (Method 2), tert-butyl(1-(3-nitropyridin-4-yl)piperidin-3-yl)methylcarbamate was reducedyielding tert-butyl(1-(3-aminopyridin-4-yl)piperidin-3-yl)methylcarbamate (72%). LCMS(m/z): 307.2 (MH⁺); LC R_(t)=2.28 min. ¹H NMR (CDCl₃): δ 8.01 (s, 1H),7.96 (d, J=5.4 Hz, 1H), 6.78 (d, J=5.4 Hz, 1H), 4.60 (bs, 1H), 3.68 (bs,2H), 3.04-3.28 (m, 4H), 2.53-2.65 (m, 1H), 2.35-2.47 (m, 1H), 1.77-1.93(m, 3H), 1.55-1.75 (m, 2H), 1.44 (s, 9H).

Example 54 Synthesis of tert-butyl1-(2-aminophenyl)piperidin-3-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(2-nitrophenyl)piperidin-3-ylcarbamate was reduced yielding tert-butyl1-(2-aminophenyl)piperidin-3-ylcarbamate, (90%). LCMS (m/z): 292.2(MH⁺). LC R_(t)=2.17 min.

Example 55 Synthesis of tert-butyl1-(2-aminophenyl)piperidin-4-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(2-nitrophenyl)piperidin-4-ylcarbamate was reduced yielding tert-butyl1-(2-aminophenyl)piperidin-4-ylcarbamate, (90%). LCMS (m/z): 292.1(MH⁺); LC R_(t)=2.13 min.

Example 56 Synthesis of tert-butyl4-(2-aminophenyl)piperazine-1-carboxylate

Following the method of Example 49 (Method 2), tert-butyl4-(2-nitrophenyl)piperazine-1-carboxylate was reduced for 2 hoursyielding tert-butyl 4-(2-aminophenyl)piperazine-1-carboxylate, (99%).LCMS (m/z): 278.2 (MH⁺); LC R_(t)=2.22 min.

Example 57 Synthesis of tert-butyl1-(3-aminopyridin-2-yl)piperidin-3-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(3-nitropyridin-2-yl)piperidin-3-ylcarbamate was reduced yieldingtert-butyl 1-(3-aminopyridin-2-yl)piperidin-3-ylcarbamate, (95%). LCMS(m/z): 293.2 (MH⁺); LC R_(t)=1.87 min.

Example 58 Synthesis of4-(4-(dimethylamino)piperidin-1-yl)pyridin-3-amine

Following the method of Example 49 (Method 2),N,N-dimethyl-1-(3-nitropyridin-4-yl)piperidin-4-amine was reducedyielding 4-(4-(dimethylamino)piperidin-1-yl)pyridin-3-amine LCMS (m/z):221.2 (MH⁺);

Example 59 Synthesis of4-(1,4-dioxa-8-azaspiro[4.5]decan-8-yl)pyridin-3-amine

Following the method of Example 49 (Method 2),8-(3-nitropyridin-4-yl)-1,4-dioxa-8-azaspiro[4.5] decane was reducedyielding 4-(1,4-dioxa-8-azaspiro[4.5]decan-8-yl)pyridin-3-amine. LCMS(m/z): 236.2 (MH⁺).

Example 60 Synthesis of tert-butyl4-(3-aminopyridin-4-yl)-1,4-diazepane-1-carboxylate

Following the method of Example 49 (Method 2), tert-butyl4-(3-nitropyridin-4-yl)-1,4-diazepane-1-carboxylate was reduced yieldingtert-butyl 4-(3-aminopyridin-4-yl)-1,4-diazepane-1-carboxylate. LCMS(m/z): 293.3 (MH⁺).

Example 61 Synthesis ofN⁴-[2-(dimethylamino)ethyl]-N⁴-methylpyridine-3,4-diamine

Following the method of Example 49 (Method 2),N¹,N¹,N²-trimethyl-N²-(3-nitropyridin-4-yl)ethane-1,2-diamine wasreduced yieldingN⁴-[2-(dimethylamino)ethyl]-N⁴-methylpyridine-3,4-diamine Concentratedand took on as is. LCMS (m/z): 195.2 (MH⁺); LC R_(t)=0.31 min.

Example 62 Synthesis of tert-butyl1-(3-aminopyridin-4-yl)pyrrolidin-3-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(3-nitropyridin-4-yl)pyrrolidin-3-ylcarbamate was reduced yieldingtert-butyl 1-(3-aminopyridin-4-yl)pyrrolidin-3-ylcarbamate. Concentratedand took on as is. LCMS (m/z): 279.1 (MH⁺); LC R_(t)=1.75 min.

Example 63 Synthesis of(R)-tert-butyl-[1-(3-aminopyridin-4-yl)pyrrolidin-2-yl]methylcarbamate)

Following the method of Example 49 (Method 2), (R)-tert-butyl[1-(3-nitropyridin-4-yl)pyrrolidin-2-yl]methyl carbamate was reducedyielding (R)-tert-butyl[1-(3-aminopyridin-4-yl)pyrrolidin-2-yl]methylcarbamate. Concentratedand took on as is. LCMS (m/z): 293.1 (MH⁺); LC R_(t)=1.79 min.

Example 64 Synthesis of 4-(piperidin-1-yl)pyrimidin-5-amine

Following the method of Example 49 (Method 2),2-chloro-5-nitro-4-(piperidin-1-yl)pyrimidine was reduced yielding4-(piperidin-1-yl)pyrimidin-5-amine as the HCl salt (100%). LCMS (m/z):179.0 (MH⁺); LC R_(t)=1.51 min

Example 65 Synthesis of tert-butyl1-(2-amino-3-fluorophenyl)piperidin-4-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(3-fluoro-2-nitrophenyl)piperidin-4-ylcarbamate was reduced in 75 minyielding tert-butyl 1-(2-amino-3-fluorophenyl)piperidin-4-yl-carbamate(95%). LCMS (m/z): 310.2 (MH⁺); LC R_(t)=2.64 min.

Example 66 Synthesis of tert-butyl1-(2-amino-5-fluorophenyl)piperidin-4-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(5-fluoro-2-nitrophenyl)piperidin-4-ylcarbamate was reduced in 75 minyielding tert-butyl 1-(2-amino-5-fluorophenyl)piperidin-4-ylcarbamate(97%). LCMS (m/z): 310.1 (MH⁺); LC R_(t)=2.25 min.

Example 67 Synthesis of tert-butyl1-(2-amino-4-fluorophenyl)piperidin-4-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(4-fluoro-2-nitrophenyl)piperidin-4-ylcarbamate was reduced yieldingtert-butyl 1-(2-amino-4-fluorophenyl)piperidin-4-ylcarbamate (90%). LCMS(m/z): 310.1 (MH⁺); LC R_(t)=2.36 min.

Example 68 Synthesis of tert-butyl1-(2-amino-4-methoxyphenyl)piperidin-3-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(4-methoxy-2-nitrophenyl)piperidin-3-ylcarbamate was reduced for 24hours yielding tert-butyl1-(2-amino-4-methoxyphenyl)piperidin-3-yl-carbamate (25%). LCMS (m/z):322.2 (MH⁺); LC R_(t)=2.27 min.

Example 69 Synthesis of tert-butyl1-(2-amino-4-methoxyphenyl)piperidin-4-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(4-methoxy-2-nitrophenyl)piperidin-4-ylcarbamate was reduced for 24hours yielding tert-butyl1-(2-amino-4-methoxyphenyl)piperidin-4-ylcarbamate (50%). LCMS (m/z):322.2 (MH⁺); LC R_(t)=2.16 min.

Example 70 Synthesis of tert-butyl4-(2-amino-4-methoxyphenyl)piperazine-1-carboxylate

Following the method of Example 49 (Method 2), tert-butyl4-(4-methoxy-2-nitrophenyl)piperazine-1-carboxylate was reduced for 24hours yielding tert-butyl4-(2-amino-4-methoxyphenyl)piperazine-1-carboxylate (20%). LCMS (m/z):308.2 (MH⁺); LC R_(t)=2.35 min.

Example 71 Synthesis of tert-butyl4-(2-amino-4-methylphenyl)piperazine-1-carboxylate

Following the method of Example 49 (Method 2), tert-butyl4-(4-methyl-2-nitrophenyl)piperazine-1-carboxylate was reduced for 2hours yielding tert-butyl4-(2-amino-4-methylphenyl)piperazine-1-carboxylate (93%). LCMS (m/z):292.1 (MH⁺); LC R_(t)=2.33 min.

Example 72 Synthesis of tert-butyl1-(2-amino-4-methylphenyl)piperidin-4-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(4-methyl-2-nitrophenyl)piperidin-4-ylcarbamate was reduced for 2hours yielding tert-butyl1-(2-amino-4-methylphenyl)piperidin-4-ylcarbamate (95%). LCMS (m/z):306.2 (MH⁺); LC R_(t)=2.22 min.

Example 73 Synthesis of tert-butyl1-(2-amino-4-methylphenyl)piperidin-3-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(4-methyl-2-nitrophenyl)piperidin-3-ylcarbamate was reduced for 2hours yielding tert-butyl1-(2-amino-4-methylphenyl)piperidin-3-ylcarbamate (95%). LCMS (m/z):306.2 (MH⁺); LC R_(t)=2.30 min.

Example 74 Synthesis of tert-butyl4-(2-amino-5-methylphenyl)piperazine-1-carboxylate

Following the method of Example 49 (Method 2), tert-butyl4-(5-methyl-2-nitrophenyl)piperazine-1-carboxylate was reduced yieldingtert-butyl 4-(2-amino-5-methylphenyl)piperazine-1-carboxylate (90%).LCMS (m/z): 292.1 (MH⁺); LC R_(t)=2.29 min.

Example 75 Synthesis of tert-butyl1-(2-amino-5-methylphenyl)piperidin-4-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(5-methyl-2-nitrophenyl)piperidin-4-ylcarbamate was reduced for 1 houryielding tert-butyl 1-(2-amino-5-methylphenyl)piperidin-4-yl-carbamate(93%). LCMS (m/z): 306.2 (MH⁺); LC R_(t)=2.25 min.

Example 76 Synthesis of tert-butyl1-(2-amino-5-methylphenyl)piperidin-3-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(5-methyl-2-nitrophenyl)piperidin-3-ylcarbamate was reduced for 1 houryielding tert-butyl 1-(2-amino-5-methylphenyl)piperidin-3-yl-carbamate(95%). LCMS (m/z): 306.2 (MH⁺); LC R_(t)=2.29 min.

Example 77 Synthesis of tert-butyl1-(2-amino-4-(trifluoromethyl)phenyl)piperidin-3-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(2-nitro-4-(trifluoromethyl)phenyl)piperidin-3-yl carbamate in MeOHwas reduced yielding tert-butyl1-(2-amino-4-(trifluoromethyl)phenyl)piperidin-3-ylcarbamate (95%). LCMS(m/z): 360.1 (MH⁺); LC R_(t)=3.30 min

Example 78 Synthesis of tert-butyl2-amino-4-(trifluoromethyl)phenyl)piperidin-4-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(2-nitro-4-(trifluoromethyl)phenyl)piperidin-4-yl-carbamate in MeOHwas reduced yielding tert-butyl1-(2-amino-4-(trifluoromethyl)phenyl)piperidin-4-ylcarbamate (97%). LCMS(m/z): 360.1 (MH⁺); LC R_(t)=3.20 min.

Example 79 Synthesis of tert-butyl4-(2-amino-4-(trifluoromethyl)phenyl)piperazine-1-carboxylate

Following the method of Example 49 (Method 2), tert-butyl4-(2-nitro-4-(trifluoro-methyl)phenyl)piperazine-1-carboxylate in MeOHwas reduced yielding tert-butyl4-(2-amino-4-(trifluoromethyl)phenyl)piperazine-1-carboxylate (99%).LCMS (m/z): 346.1 (MH⁺); LC R_(t)=3.38 min.

Example 80 Synthesis of tert-butyl1-(2-amino-4-cyanophenyl)piperidin-3-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(4-cyano-2-nitrophenyl)piperidin-3-ylcarbamate was reduced yieldingtert-butyl 1-(2-amino-4-cyanophenyl)piperidin-3-ylcarbamate (95%). LCMS(m/z): 317.2 (MH⁺); LC R_(t)=2.92 min.

Example 81 Synthesis of tert-butyl1-(2-amino-4-(1H-pyrazol-5-yl)phenyl)piperidin-4-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(2-nitro-4-(1H-pyrazol-5-yl)phenyl)piperidin-4-ylcarbamate was reducedyielding tert-butyl1-(2-amino-4-(1H-pyrazol-5-yl)phenyl)piperidin-4-ylcarbamate (87%). LCMS(m/z): 258.1 (MH⁺); LC R_(t)=2.15 min.

Example 82 Synthesis of tert-butyl1-(2-amino-4-(methylsulfonyl)phenyl)piperidin-4-ylcarbamate

Following the method of Example 49 (Method 2), tert-butyl1-(4-(methylsulfonyl)-2-nitrophenyl)piperidin-4-yl-carbamate was reducedyielding tert-butyl1-(2-amino-4-(methylsulfonyl)phenyl)piperidin-4-ylcarbamate (76%). LCMS(m/z): 370.1 (MH⁺); LC R_(t)=2.52 min.

Example 83 Synthesis of tert-butyl2-(3-aminopyridin-4-yloxy)ethylcarbamate

Following the method of Example 49 (Method 2), tert-butyl2-(3-nitropyridin-4-yloxy)ethylcarbamate was reduced to yield tert-butyl2-(3-aminopyridin-4-yloxy)ethylcarbamate. LCMS (m/z): 254.1 (MH⁺); LCR_(t)=1.76 min.

Method 3 Example 84 Synthesis of tert-butyl3-(3-nitropyridin-4-yl)piperazin-1-yl)-3-oxopropylcarbamate

A solution containing 1.0 eq each of 1-(3-nitropyridin-4-yl)piperazine,N-Boc-beta-alanine, HOAT and EDC in DCM, at a concentration of 0.1 M,was stirred for 16 hours. The solution was diluted with EtOAc and waswashed with H₂O, Na₂CO_(3 (sat.)), NaCl_((sat)), was dried over MgSO₄,was filtered and the volatiles were removed in vacuo yielding tert-butyl3-(4-(3-nitropyridin-4-yl)piperazin-1-yl)-3-oxopropylcarbamate (93%).LCMS (m/z): 379.9 (MH⁺); LC R_(t)=1.92 min.

Example 85 Synthesis of tert-butyl3-(4-(3-aminopyridin-4-yl)piperazin-1-yl)-3-oxopropylcarbamate

Following the method of Example 49 (Method 2), tert-butyl3-(4-(3-nitropyridin-4-yl)piperazin-1-yl)-3-oxopropylcarbamate wasreduced yielding tert-butyl(tert-butyl3-(4-(3-aminopyridin-4-yl)piperazin-1-yl)-3-oxopropylcarbamate (99%yield). LCMS (m/z): 349.9 (MH⁺); LC R_(t)=1.84 min.

Example 86 Synthesis of tert-butyl2-(4-(3-nitropyridin-4-yl)piperazin-1-yl)-2-oxoethylcarbamate

Following the method of Example 84 (Method 3),1-(3-nitropyridin-4-yl)piperazine was coupled to N-Boc-glycine yieldingtert-butyl 2-(4-(3-nitropyridin-4-yl)piperazin-1-yl)-2-oxoethylcarbamate(99% yield). LCMS (m/z): 365.8 (MH⁺); LC R_(t)=1.81 min

Example 87 Synthesis of tert-butyl2-(4-(3-aminopyridin-4-yl)piperazin-1-yl)-2-oxoethylcarbamate

Following the method of Example 49 (Method 2), tert-butyl24443-nitropyridin-4-yl)piperazin-1-yl)-2-oxoethylcarbamate was reducedyielding tert-butyl(tert-butyl2-(4-(3-aminopyridin-4-yl)piperazin-1-yl)-2-oxoethylcarbamate (88%yield). LCMS (m/z): 335.8 (MH⁺); LC R_(t)=1.79 min.

Method 4 Example 88 Synthesis of 4-nitro-3-(piperidin-1-yl)pyridine1-oxide

3-bromo-4-nitropyridine-N-oxide (1.0 equiv.) and piperidine (2.0 equiv.)in ethanol, at a concentration of 0.2 M, was heated at reflux for 16hours. Upon cooling the ethanol was removed in vacuo. The residue waspartitioned between EtOAc and Na₂CO_(3 (sat.)), and washed further withH₂O, NaCl_((sat)), was dried over MgSO₄, was filtered and the volatileswere removed in vacuo yielding 4-nitro-3-(piperidin-1-yl)pyridine1-oxide (92%). LCMS (m/z): 224.0 (MH⁺); LC R_(t)=2.48 min.

Example 89 Synthesis of tert-butyl1-(4-nitropyridin-3-yl)piperidin-3-ylcarbamate

The method of Example 88 (Method 4) was followed using 1 eq each of3-bromo-4-nitropyridine-N-oxide, 3-N-Boc-amino piperidine anddiisopropylethylamine yielding tert-butyl1-(4-nitropyridin-3-yl)piperidin-3-ylcarbamate (65%). LCMS (m/z): 339.1(MH⁺); LC R_(t)=2.88 min.

Method 5 Example 90 Synthesis of 3-(piperidin-1-yl)pyridin-4-amine

To a solution of 4-nitro-3-(piperidin-1-yl)pyridine 1-oxide (1.0 equiv.)in ethanol, at a concentration of 0.1 M, was added 10% palladium oncarbon (0.1 eq.). The resultant heterogeneous solution was put under anatmosphere of hydrogen and was stirred for 15 hours. At this time LC/MSanalysis indicated that the nitro was reduced to the amine, but theN-oxide was remaining. More 10% palladium on carbon (0.2 eq.) was addedand the mixture was resubmitted to a balloon atmosphere of hydrogen.After stirring for 24 hours, more 10% palladium on carbon (0.2 eq.) wasadded and the mixture was resubmitted to a balloon atmosphere ofhydrogen. After stirring for an additional 3 days the mixture wasfiltered through a pad of celite eluting with methanol. The volatileswere removed in vacuo yielding 3-(piperidin-1-yl)pyridin-4-amine (73%).LCMS (m/z): 178.0 (MH⁺); LC R_(t)=1.66 min

Example 91 Synthesis of tert-butyl1-(4-aminopyridin-3-yl)piperidin-3-ylcarbamate

The method of Example 90 (Method 5) was followed using 1 eq oftert-butyl 1-(4-nitropyridin-3-yl)piperidin-3-ylcarbamate in 1:1ethanol/ethyl acetate at 30 psi for 72 hours, yielding tert-butyl1-(4-aminopyridin-3-yl)piperidin-3-ylcarbamate (79%). LCMS (m/z): 293.1(MH⁺); LC R_(t)=2.14 min.

Method 6 Example 92 Synthesis of 4-cyclohexenyl-3-nitropyridine

A solution of 4-chloro-3-nitro pyridine (1 eq.), cyclohexenyl boronicacid (1.7 eq.), and Pd(dppf)Cl₂—CH₂Cl₂ (0.05 eq) in 3:1 DME/2M Na₂CO₃,at a concentration of 0.1 M was heated at 95° C. for 16 hours. Uponcooling the reaction was partitioned between EtOAc and H₂O, was washedwith NaCl_((sat)), dried over MgSO₄, was filtered and the volatiles wereremoved in vacuo. The material was purified by SiO₂ chromatography (20%EtOAc/hexanes eluant) to yield 4-cyclohexenyl-3-nitropyridine

(82%). LCMS (m/z): 205.0 (MH⁺); LC R_(t)=3.84 min.

Example 93 Synthesis of 3-nitro-4-o-tolylpyridine

The method of Example 92 (Method 6) was followed using ortho-tolylboronic acid for 3 hours, yielding 3-nitro-4-o-tolylpyridine (88%). LCMS(m/z): 215.1 (MH⁺); LC R_(t)=3.58 min.

Method 7 Example 94 Synthesis of 4-cyclohexenylpyridin-3-amine

A heterogeneous solution of 4-cyclohexenyl-3-nitropyridine (1.0 eq.) andiron (6.0 eq) in acetic acid, at a concentration of 0.4 M, was stirredvigorously for 2 hours. The mixture was then passed through a celitepad, eluting with MeOH. Upon removal of the volatiles in vacuo, theresidue was dissolved in EtOAc, washed with Na₂CO_(3 (sat)),NaCl_((sat)), was dried over MgSO₄, was filtered and the volatiles wereremoved in vacuo yielding 4-cyclohexenylpyridin-3-amine (99%) as an oil.LCMS (m/z): 175.0 (MH⁺); LC R_(t)=1.86 min.

Example 95 Synthesis of 4-o-tolylpyridin-3-amine

The method of Example 94 (Method 7) was followed using 3-nitro-4-O—tolylpyridine yielding 4-o-tolylpyridin-3-amine (97%). LCMS (m/z): 185.1(MH⁺); LC R_(t)=1.78 min.

Example 96 Synthesis of tert-butyl1-(2-amino-4-benzoylphenyl)piperidin-3-ylcarbamate

Following method 7, tert-butyl1-(4-benzoyl-2-nitrophenyl)piperidin-3-ylcarbamate was reduced for 16hours, filtered, and concentrated. Water was added to the residue. Theresulting solids were collected by filtration and dried in vacuoyielding tert-butyl 1-(2-amino-4-benzoylphenyl)piperidin-3-ylcarbamate(90%). LCMS (m/z): 396.2 (MH⁺); LC R_(t)=3.07 min.

Example 97 Synthesis of tert-butyl1-(2-amino-4-benzoylphenyl)piperidin-4-ylcarbamate

Following method 7, tert-butyl1-(4-benzoyl-2-nitrophenyl)piperidin-4-ylcarbamate was reduced for 16hours, filtered, and concentrated. Water was added to the residue. Theresulting solids were collected by filtration and dried in vacuoyielding tert-butyl 1-(2-amino-4-benzoylphenyl)piperidin-4-ylcarbamate(83%). LCMS (m/z): 396.2 (MH⁺); LC R_(t)=2.81 min.

Example 98 Synthesis of tert-butyl4-(2-amino-4-benzoylphenyl)piperazine-1-carboxylate

Following method 7, tert-butyl1-(4-benzoyl-2-nitrophenyl)piperidin-4-ylcarbamate was reduced for 16hours, filtered, and concentrated. Water was added to the residue. Theresulting solids were collected by filtration and dried in vacuoyielding tert-butyl 4-(2-amino-4-benzoylphenyl)piperazine-1-carboxylate(61%). LCMS (m/z): 382.2 (MH⁺); LC R_(t)=3.01 min.

Example 99 Synthesis of tert-butyl 4-(4-acetyl-2-aminophenyl)piperazine-1-carboxylate

Following method 7, tert-butyl4-(4-acetyl-2-nitrophenyl)piperazine-1-carboxylate was reduced,filtered, and concentrated. Water was added to the residue. Theresulting solids were collected by filtration and dried in vacuoyielding tert-butyl 4-(4-acetyl-2-amino phenyl)piperazine-1-carboxylate(87%). LCMS (m/z): 320.2 (MH⁺); LC R_(t)=2.58 min.

Example 100 Synthesis of tert-butyl1-(4-acetyl-2-aminophenyl)piperidin-4-ylcarbamate

Following method 7, tert-butyl1-(4-acetyl-2-nitrophenyl)piperidin-4-ylcarbamate was reduced, filtered,and concentrated. Water was added to the residue. The resulting solidswere collected by filtration and dried in vacuo yielding tert-butyl1-(4-acetyl-2-amino phenyl)piperidin-4-ylcarbamate (90%). LCMS (m/z):334.2 (MH⁺); LC R_(t)=2.42 min.

Example 101 Synthesis of tert-butyl1-(4-acetyl-2-aminophenyl)piperidin-3-ylcarbamate

Following method 7, tert-butyl 1-(4-acetyl-2-nitrophenyl)piperidin-3-ylcarbamate was reduced, filtered, and concentrated.Water was added to the residue. The resulting solids were collected byfiltration and dried in vacuo yielding tert-butyl 1-(4-acetyl-2-aminophenyl)piperidin-3-ylcarbamate (88%). LCMS (m/z): 334.2 (MH⁺); LCR_(t)=2.49 min.

Example 102 Synthesis of tert-butyl4-(2-amino-4-chlorophenyl)piperazine-1-carboxylate

Following method 7, tert-butyl4-(4-chloro-2-nitrophenyl)piperazine-1-carboxylate was reduced,filtered, and concentrated. Water was added to the residue. Theresulting solids were collected by filtration and dried in vacuoyielding tert-butyl 4-(2-amino-4-chloro phenyl)piperazine-1-carboxylate(80%). LCMS (m/z): 312.1 (MH⁺); LC R_(t)=2.85 min.

Example 103 Synthesis of tert-butyl1-(2-amino-4-chlorophenyl)piperidin-4-ylcarbamate

Following method 7, tert-butyl1-(4-chloro-2-nitrophenyl)piperidin-4-ylcarbamate was reduced, filtered,and concentrated. Water was added to the residue. The resulting solidswere collected by filtration and dried in vacuo yielding tert-butyl1-(2-amino-4-chloro phenyl)piperidin-4-ylcarbamate (68%). LCMS (m/z):326.1 (MH⁺); LC R_(t)=2.67 min.

Example 104 Synthesis of tert-butyl1-(2-amino-4-chlorophenyl)piperidin-3-ylcarbamate

Following method 7, tert-butyl1-(4-chloro-2-nitrophenyl)piperidin-3-ylcarbamate was reduced, filtered,and concentrated. Water was added to the residue. The resulting solidswere collected by filtration and dried in vacuo yielding tert-butyl1-(2-amino-4-chloro phenyl)piperidin-3-ylcarbamate (85%). LCMS (m/z):326.1 (MH⁺); LC R_(t)=2.76 min.

Example 105 Synthesis of tert-butyl4-(4-(cyclopropanecarbonyl)-2-nitrophenyl)piperazine-1-carboxylate

Following method 7, tert-butyl4-(4-(cyclopropanecarbonyl)-2-nitrophenyl)piperazine-1-carboxylate wasreduced, filtered, and concentrated. Water was added to the residue. Theresulting solids were collected by filtration and dried in vacuoyielding tert-butyl 1-(2-amino-4-chloro phenyl)piperidin-4-ylcarbamate(90%). LCMS (m/z): 346.2 (MH⁺); LC R_(t)=2.83 min.

Example 106 Synthesis of tert-butyl2-amino-4-(cyclopropanecarbonyl)phenyl)piperidin-4-ylcarbamate

Following method 7, tert-butyl1-(4-(cyclopropanecarbonyl)-2-nitrophenyl)piperidin-4-ylcarbamate wasreduced, filtered, and concentrated. Water was added to the residue. Theresulting solids were collected by filtration and dried in vacuoyielding tert-butyl1-(2-amino-4-(cyclopropanecarbonyl)phenyl)piperidin-4-ylcarbamate (93%).LCMS (m/z): 360.1 (MH⁺); LC R_(t)=2.65 min.

Example 107 Synthesis of tert-butyl2-amino-4-(cyclopropanecarbonyl)phenyl)piperidin-3-ylcarbamate

Following method 7, tert-butyl1-(4-(cyclopropanecarbonyl)-2-nitrophenyl)piperidin-3-ylcarbamate wasreduced, filtered, and concentrated. Water was added to the residue. Theresulting solids were collected by filtration and dried in vacuoyielding tert-butyl1-(2-amino-4-(cyclopropanecarbonyl)phenyl)piperidin-3-ylcarbamate (90%).LCMS (m/z): 360.1 (MH⁺); LC R_(t)=2.74 min.

Method 8 Example 108 Synthesis of6-amino-5-(4-(trifluoromethyl)piperidin-1-yl)pyrimidin-2(1H)-one

A solution of 5-bromocytosine (1.0 equiv.),4-(trifluoromethyl)piperidine (1.25 equiv.) and diisopropylethylamine(1.25 equiv.) in N-methylpyrrolidinone (NMP), at a concentration of0.525 M, was degassed by bubbling argon through for 10 minutes in a 125mL high-pressure glass vessel. The glass bomb was then sealed and heatedat 120° C. for 3 days. The reaction mixture was purified directly byreverse-phase HPLC and lyophilized yielding a TFA salt of the product asa crunchy orange solid (50%). LCMS (m/z): 263.0 (MH⁺); LC R_(t)=1.81min.

Example 109 Synthesis of6-amino-5-(4,4-difluoropiperidin-1-yl)pyrimidin-2(1H)-one

Method 8 was followed using 1 eq. of 5-bromocytosine, 1.25 eq. of3-fluoropiperidine and 2.5 eq. of diisopropylethylamine at 120° C. for 2days yielding 6-amino-5-(4,4-difluoropiperidin-1-yl)pyrimidin-2(1H)-oneas an orange crunchy solid (34%). LCMS (m/z): 231.0 (MH⁺); LC R_(t)=1.28min.

Example 110 Synthesis of6-amino-5-(3-fluoropiperidin-1-yl)pyrimidin-2(1H)-one

Method 8 was followed using 1 eq. of 5-bromocytosine, 1.25 eq. of3-fluoropiperidine and 2.5 eq. of diisopropylethylamine at 120° C. for 2days yielding 6-amino-5-(3-fluoropiperidin-1-yl)pyrimidin-2(1H)-one asan orange crunchy solid (24%). LCMS (m/z): 213.0 (MH⁺); LC R_(t)=1.07min.

Example 111 Synthesis oftert-butyl(1-(6-amino-2-oxo-1,2-dihydropyrimidin-5-yl)piperidin-3-yl)methylcarbamate

Method 8 was followed using 1 eq. of 5-bromocytosine, 1.05 eq. oftert-butyl piperidin-3-ylmethylcarbamate and 1.05 eq. ofdiisopropylethylamine yieldingtert-butyl(1-(6-amino-2-oxo-1,2-dihydropyrimidin-5-yl)piperidin-3-yl)methylcarbamateas an orange crunchy solid (18%). LCMS (m/z): 324.1 (MH⁺); LC R_(t)=1.90min.

Example 112 Synthesis of tert-butyl(1-(6-amino-2-oxo-1,2-dihydropyrimidin-5-yl)piperidin-3-yl)carbamate

Method 8 was followed using 1 eq. of 5-bromocytosine, 1.05 eq. oftert-butyl piperidin-3-ylmethylcarbamate and 1.05 eq. ofdiisopropylethylamine yieldingtert-butyl(1-(6-amino-2-oxo-1,2-dihydropyrimidin-5-yl)piperidin-3-yl)carbamateas an orange crunchy solid (26%). LCMS (m/z): 310.1 (MH⁺); LC R_(t)=1.78min.

Example 113 Synthesis of tert-butyl3-(4-(6-amino-2-oxo-1,2-dihydropyrimidin-5-yl)piperazin-1-yl)-3-oxopropylcarbamate

Method 8 was followed using 1 eq. of 5-bromocytosine, 1.5 eq. often-butyl 3-oxo-3-(piperazin-1-yl)propylcarbamate and 1.2 eq. ofdiisopropylethylamine yielding tert-butyl3-(4-(6-amino-2-oxo-1,2-dihydropyrimidin-5-yl)piperazin-1-yl)-3-oxopropylcarbamate as an orange crunchy solid (65%). LCMS (m/z): 367.2 (MH⁺); LCR_(t)=1.68 min.

Example 114 Synthesis of 6-amino-5-(piperidin-1-yl)pyrimidin-2(1H)-one

Method 8 was followed using 1 eq. of 5-bromocytosine and 15 eq. ofpiperidine (as solvent). The reaction was cooled and added to CH₂Cl₂ andH₂O. The solid was filtered, rinsed with H₂O, and dried yielding6-amino-5-(piperidin-1-yl)pyrimidin-2(1H)-one as a solid (89%). LCMS(m/z): 195.0 (MH⁺); LC R_(t)=1.28 min.

Method 9 Example 115 Synthesis of3-amino-N-(4-(piperidin-1-yl)pyridin-3-yl)pyrazine-2-carboxamide

A solution of 1 eq of 4-(piperidin-1-yl)pyridin-3-amine and 2 eq each of3-aminopyrazine-2-carboxylic acid, HOAT and EDC in NMP, at aconcentration of 0.2 M, was stirred for 48 hours at which time themixture was directly purified by HPLC. Upon lyophilization, the TFA saltof 3-amino-N-(4-(piperidin-1-yl)pyridin-3-yl)pyrazine-2-carboxamide wasobtained (61%). Alternatively, the HPLC fractions could be added toEtOAc and solid Na₂CO₃, separated and washed with NaCl_((sat)) Upondrying over MgSO₄, filtering and removing the volatiles in vacuo thefree base was obtained. Upon dissolving in MeCN/H₂O, adding 1 eq. of 1 NHCl and lyophilizing, the HCl salt of3-amino-N-(4-(piperidin-1-yl)pyridin-3-yl)pyrazine-2-carboxamide wasobtained (40%). LCMS (m/z): 298.8 (MH⁺); LC R_(t)=1.88 min. ¹H NMR ofHCl salt (DMSO_(d-6)): δ 10.45 (s, 1H), 8.55 (d, J=0.9, 1H), 8.32 (d,J=2.1, 1H), 8.27 (dd, J=5.7, 1H), 7.93 (d, J=1.8, 1H), 7.57 (s, 1H),7.32 (d, J=6.9, 1H), 3.76 (s, 4H), 1.59 (s, 6H).

Example 116 Synthesis of3-amino-6-bromo-N-(4-(piperidin-1-yl)pyridin-3-yl)picolinamide

The method of Example 115 (Method 9) was followed using4-(piperidin-1-yl)pyridin-3-amine yielding3-amino-6-bromo-N-(4-(piperidin-1-yl)pyridin-3-yl)picolinamide (32%).LCMS (m/z): 376.1 (MH⁺); LC R_(t)=2.77 min.

Example 117 Synthesis of3-amino-6-bromo-N-(4-o-tolylpyridin-3-yl)picolinamide

The method of Example 115 (Method 9) was followed using 4-O—tolylpyridin-3-amine yielding3-amino-6-bromo-N-(4-o-tolylpyridin-3-yl)picolinamide (74%). LCMS (m/z):383.0 (MH⁺); LC R_(t)=2.99 min.

The following compounds were prepared using Method 9.

Example Structure Name MH+ LC 118

3-amino-N-(4-cyclohex-1-en- 1-ylpyridin-3-yl)pyrazine-2- carboxamide296.1 2.32 119

3-amino-6-bromo-N-(4- piperidin-1-ylpyridin-3- yl)pyrazine-2-carboxamide377 2.46 120

3,5-diamino-6-chloro-N-(3- piperidin-1-ylpyridin-4-yl)-pyrazine-2-carboxamide 348.1 2.43 121

3,5-diamino-6-chloro-N-(4- piperidin-1-ylpyridin-3-yl)-pyrazine-2-carboxamide 347.8 2.17 122

3-amino-N-(3-piperidin-1-yl- pyridin-4-yl)pyrazine-2-carbox- amide 298.82.26 123

3-amino-N-(4-piperidin-1-yl- pyridin-3-yl)pyrazine-2-carbox- amide 298.81.88 124

3-amino-6-bromo-N-(4-{[2- (dimethylamino)ethyl]-(methyl)amino}pyridin-3- yl)pyrazine-2-carboxamide 396.0 125

3-amino-N-(4-chloropyridin-3- yl)pyrazine-2-carboxamide 250 1.34 126

3-amino-N-{4-[4-(dimethyl- amino)piperidin-1-yl]pyridin-3-yl}pyrazine-2-carboxamide 342.2 0.92 127

3-amino-N-[4-(1,4-dioxa-8- azaspiro[4.5]dec-8-yl)pyridin-3-yl]pyrazine-2-carboxamide 357.1 1.66 128

3-amino-N-(4-piperidin-1-yl- pyrimidin-5-yl)pyrazine-2- carboxamide299.7 1.72 129

3-amino-N-(5-carbamoyl-2- piperidin-1-ylphenyl)pyrazine-2- carboxamide340.8 3.11 130

3-amino-N-(2-oxo-5-piperidin-1- yl-2,3-dihydropyrimidin-4-yl)-pyrazine-2-carboxamide 315.7 2.21 131

3,5-diamino-6-chloro-N-(2-oxo- 5-piperidin-1-yl-2,3-dihydro-pyrimidin-4-yl)pyrazine-2- carboxamide 365.1 2.2 132

3-amino-6-bromo-N-(2-oxo-5- piperidin-1-yl-2,3-dihydro-pyrimidin-4-yl)pyrazine-2- carboxamide 394 2.81 133

3-amino-6-bromo-N-(5- carbamoyl-2-piperidin-1-yl-phenyl)pyrazine-2-carboxamide 419 3.98 134

3,5-diamino-N-[5-(3-amino- piperidin-1-yl)-2-oxo-2,3-dihydropyrimidin-4-yl]-6- chloropyrazine-2-carboxamide 380.1 1.29 135

3-amino-N-{5-[3-(amino- methyl)piperidin-1-yl]-2-oxo-2,3-dihydropyrimidin-4-yl}- pyrazine-2-carboxamide 345.1 1.39 136

3-amino-N-[5-(2-furyl)-2-oxo- 2,3-dihydropyrimidin-4-yl]-pyrazine-2-carboxamide 299 1.93 137

3-amino-N-[2-oxo-5-(2-thienyl)- 2,3-dihydropyrimidin-4-yl]-pyrazine-2-carboxamide 315 2.02 138

3-amino-N-[2-oxo-5-(1,3- thiazol-2-yl)-2,3-dihydro-pyrimidin-4-yl]pyrazine-2- carboxamide 316 1.92 139

3-amino-N-[5-(1-methyl-1H- pyrrol-2-yl)-2-oxo-2,3-dihydro-pyrimidin-4-yl]pyrazine-2- carboxamide 312.1 2.02 140

3-amino-N-(2-oxo-5-pyridin-2- yl-2,3-dihydropyrimidin-4-yl)-pyrazine-2-carboxamide 310 1.82 141

3-amino-6-bromo-N-[5-(2- furyl)-2-oxo-2,3-dihydro-pyrimidin-4-yl]pyrazine-2- carboxamide 376.9 3.98 142

3-amino-6-bromo-N-[2-oxo-5- (2-thienyl)-2,3-dihydro-pyrimidin-4-yl]pyrazine-2- carboxamide 392.9 2.66 143

3-amino-6-bromo-N-[2-oxo-5- (1,3-thiazol-2-yl)-2,3-dihydro-pyrimidin-4-yl]pyrazine-2- carboxamide 393.9 2.53 144

3-amino-6-bromo-N-[5-(1- methyl-1H-pyrrol-2-yl)-2-oxo-2,3-dihydropyrimidin-4-yl]- pyrazine-2-carboxamide 390 2.57 145

3-amino-6-bromo-N-(2-oxo-5- pyridin-2-yl-2,3-dihydro-pyrimidin-4-yl)pyrazine-2- carboxamide 387.9 2.36 146

3-amino-N-{2-oxo-5-[3- (trifluoromethyl)piperidin-1-yl]-2,3-dihydropyrimidin-4-yl}- pyrazine-2-carboxamide 384.1 2.59 147

3-amino-6-bromo-N-{2-oxo-5- [3-(trifluoromethyl)piperidin-1-yl]-2,3-dihydropyrimidin-4-yl}- pyrazine-2-carboxamide 462 3.12 148

3,5-diamino-6-chloro-N-{2-oxo- 5-[3-(trifluoromethyl)piperidin-1-yl]-2,3-dihydropyrimidin-4- yl}pyrazine-2-carboxamide 433.1 2.51 149

3-amino-N-[5-(4-fluoro- piperidin-1-yl)-2-oxo-2,3-dihydropyrimidin-4-yl]pyrazine- 2-carboxamide 334.1 2 150

3-amino-6-bromo-N-[5-(4- fluoropiperidin-1-yl)-2-oxo-2,3-dihydropyrimidin-4-yl]pyrazine- 2-carboxamide 412 2.58 151

3,5-diamino-6-chloro-N-[5-(4- fluoropiperidin-1-yl)-2-oxo-2,3-dihydropyrimidin-4-yl]pyrazine- 2-carboxamide 383.1 2.02 152

3-amino-N-{2-oxo-5-[4- (trifluoromethyl)piperidin-1-yl]-2,3-dihydropyrimidin-4-yl}- pyrazine-2-carboxamide 384.1 2.62 153

3-amino-6-bromo-N-{2-oxo-5- [4-(trifluoromethyl)piperidin-1-yl]-2,3-dihydropyrimidin-4-yl}- pyrazine-2-carboxamide 462 3.08 154

3-amino-N-[5-(3-fluoro- piperidin-1-yl)-2-oxo-2,3-dihydropyrimidin-4-yl]pyrazine- 2-carboxamide 334.1 1.99 155

3-amino-6-bromo-N-[5-(3- fluoropiperidin-1-yl)-2-oxo-2,3-dihydropyrimidin-4-yl]pyrazine- 2-carboxamide 412 2.58 156

3-amino-N-[5-(4,4-difluoro- piperidin-1-yl)-2-oxo-2,3-dihydropyrimidin-4-yl]pyrazine- 2-carboxamide 352.1 2.21 157

3-amino-6-bromo-N-[5-(4,4- difluoropiperidin-1-yl)-2-oxo-2,3-dihydropyrimidin-4-yl]- pyrazine-2-carboxamide 430 2.78 158

3-amino-N-(2-oxo-5-piperidin-1- yl-2,3-dihydropyrimidin-4-yl)-pyridine-2-carboxamide 315.1 1.97 159

3,5-diamino-6-chloro-N-{2-oxo- 5-[4-(trifluoromethyl)piperidin-1-yl]-2,3-dihydropyrimidin-4- yl}pyrazine-2-carboxamide 433.1 2.5 160

3,5-diamino-6-chloro-N-[5-(3- fluoropiperidin-1-yl)-2-oxo-2,3-dihydropyrimidin-4-yl]pyrazine- 2-carboxamide 383.1 1.99 161

3,5-diamino-6-chloro-N-[5-(4,4- difluoropiperidin-1-yl)-2-oxo-2,3-dihydropyrimidin-4-yl]- pyrazine-2-carboxamide 401.1 2.23 162

3-Amino-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (3-hydroxy-3-methyl-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide422.2 2.77 163

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid (3- hydroxy-3-trifluoromethyl- 3,4,5,6-tetrahydro-2H-[1,4′]bi-pyridinyl-3′-yl)-amide 561.2 3.06 164

3-Amino-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (3-hydroxy-3-trifluoromethyl- 3,4,5,6-tetrahydro-2H-[1,4′]bi-pyridinyl-3′-yl)-amide 476.1 3.14 165

5-Amino-2-(2,6-difluoro- phenyl)-pyrimidine-4-carboxylic acid(3-hydroxy-3- trifluoromethyl-3,4,5,6-tetra-hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 495.1 2.64 166

5-Amino-2-(2-fluoro-phenyl)- pyrimidine-4-carboxylic acid (3-hydroxy-3-trifluoromethyl- 3,4,5,6-tetrahydro-2H-[1,4′]bi-pyridinyl-3′-yl)-amide 477.1 2.72 167

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid(4-azocan-1-yl-pyridin-3- yl)-amide 438.1 3.22 168

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid(4-[1,4]oxazepan-4-yl- pyridin-3-yl)-amide 425.9 2.65 169

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid(4-azepan-1-yl-pyridin-3- yl)-amide 424.1 3.14

Example 1703-amino-N-(5-carbamoyl-2-(piperidin-1-yl)phenyl)pyrazine-2-carboxamide

A solution of 3-amino-4-(piperidin-1-yl)benzamide (1.0 equiv.), HOAT(1.3 equiv.) and EDC (1.3 equiv.) in NMP at a concentration of 0.182 M,was stirred for 15 hours, then purified directly by reverse-phase HPLCand lyophilized yielding a TFA salt of3-amino-N-(5-carbamoyl-2-(piperidin-1-yl)phenyl)pyrazine-2-carboxamideas a tan powder (82%). LCMS (m/z): 341.1 (MH⁺); LC R_(t)=3.10 min.

Example 171 Synthesis of3-amino-N-(5-cyano-2-(piperidin-1-yl)phenyl)pyrazine-2-carboxamide

A milky yellow suspension of3-amino-N-(5-carbamoyl-2-(piperidin-1-yl)phenyl)pyrazine-2-carboxamide(1 equiv.) in dichloromethane (0.0247 M) was cooled in ice bath. Asolution of triflic anhydride (4.4 equiv.) in dichloromethane (0.0405 M)was added dropwise, keeping the internal temperature of the solution<2.5° C. After 5 minutes the reaction was quenched with 6 mL water andthe solution allowed to warm up to room temperature, then extracted withdichloromethane. The organics were washed with Na₂CO₃ (sat.), thenbrine, dried over Na₂SO₄, filtered and concentrated in vacuo. The cruderesidue was purified by reverse-phase HPLC and lyophilized yielding aTFA salt of3-amino-N-(5-cyano-2-(piperidin-1-yl)phenyl)pyrazine-2-carboxamide as ayellow fluffy solid (24%). LCMS (m/z): 323.1 (MH⁺); LC R_(t)=4.62 min.

Method 10 Example 172 Synthesis of3-amino-N-(4-(3-aminopiperidin-1-yl)pyridin-3-yl)pyrazine-2-carboxamide

Following method 9,3-aminopyrazine-2-carboxylic acid was coupled totert-butyl 1-(3-aminopyridin-4-yl)piperidin-3-ylcarbamate yieldingtert-butyl1-(3-(3-aminopyrazine-2-carboxamido)pyridin-4-yl)piperidin-3-ylcarbamateas the TFA salt after lyophilization of the HPLC product fraction.Alternatively, the free base could be obtained as described in method 8,(83% yield). LCMS (m/z): 414.2 (MH⁺); LC R_(t)=2.18 min.

A homogeneous solution of tert-butyl1-(3-(3-aminopyrazine-2-carboxamido)pyridin-4-yl)piperidin-3-ylcarbamate,either TFA salt or free base, in 25% TFA/DCM was allowed to sit for 2hours. Upon removal of volatiles in vacuo, the residue was purified byHPLC. Direct lyophilization led to the isolation of3-amino-N-(4-(3-aminopiperidin-1-yl)pyridin-3-yl)pyrazine-2-carboxamideas the TFA salt. Alternatively, the free base and HCl salt could beobtained as described in Method 8. LCMS (m/z): 314.1 (MH⁺); LCR_(t)=1.02 min.

An alternative manner of removing the Boc protecting group and isolatingthe HCl salt was as follows: a heterogeneous solution of tert-butyl1-(3-(3-aminopyrazine-2-carboxamido)pyridin-4-yl)piperidin-3-ylcarbamatein 4 M HCl/dioxane, at a concentration of 0.01 M, was stirred for 24hours at which time the volatiles were removed in vacuo. Aftertriturating and rinsing with diethyl ether the resultant solid wasdissolved in MeCN/H₂O and lyophilized yielding3-amino-N-(4-(3-aminopiperidin-1-yl)pyridin-3-yl)pyrazine-2-carboxamideas the HCl salt.

The following compounds were prepared using Method 10.

Example Structure Name MH+ LC 173

3-amino-N-[3-(3-aminopiperidin- 1-yl)pyridin-4-yl]pyrazine-2-carboxamide 314.1 1.03 174

3-amino-N-[3-(3-aminopiperidin- 1-yl)pyridin-4-yl]pyridine-2-carboxamide 313.2 1.23 175

3-amino-N-{4-[(3S)-3-amino- piperidin-1-yl]pyridin-3-yl}-pyridine-2-carboxamide 313.1 1.25 176

N-{4-[(3S)-3-aminopiperidin-1- yl]pyridin-3-yl}pyrazine-2- carboxamide314.1 1.05 177

3-amino-N-{4-[(3R)-3-amino- piperidin-1-yl]pyridin-3-yl}pyrazine-2-carboxamide 314.1 1.06 178

3-amino-N-{4-[4-(3-amino- propanoyl)piperazin-1-yl]-pyridin-3-yl}-6-bromopyrazine- 2-carboxamide 449.1 1.31 179

3-amino-N-{4-[4-(aminoacetyl)- piperazin-1-yl]pyridin-3-yl}-6-bromopyrazine-2-carboxamide 435.1 1.23 180

3-amino-N-{4-[3-(amino- methyl)piperidin-1-yl]pyridin-3-yl}-6-bromopyrazine-2- carboxamide 406 1.48 181

3-amino-N-[4-(3-aminopiperidin- 1-yl)pyridin-3-yl]-6-bromopyrazine-2-carboxamide 392 1.39 182

3,5-diamino-N-{4-[3-(amino- methyl)piperidin-1-yl]pyridin-3-yl}-6-chloropyrazine-2- carboxamide 377.1 1.21 183

3,5-diamino-N-[4-(3-amino- piperidin-1-yl)pyridin-3-yl]-6-chloropyrazine-2-carboxamide 363.1 1.13 184

3-amino-N-{4-[3-(aminomethyl)- piperidin-1-yl]pyridin-3-yl}pyrazine-2-carboxamide 328.1 1.13 185

3,5-Diamino-6-chloro-pyrazine- 2-carboxylic acid {4-[4-(3-amino-propionyl)-piperazin-1- yl]-pyridin-3-yl}-amide 420.1 1.2 186

3,5-diamino-N-{4-[4-(amino- acetyl)piperazin-1-yl]pyridin-3-yl}-6-chloropyrazine-2- carboxamide 406.1 1.15 187

3-amino-N-[4-(3-aminopiperidin- 1-yl)pyridin-3-yl]pyrazine-2-carboxamide 313.8 1.04 188

3-amino-N-{4-[4-(3- aminopropanoyl)piperazin-1-yl]pyridin-3-yl}pyrazine-2- carboxamide 371.1 0.88 189

3-amino-N-{4-[4-(amino- acetyl)piperazin-1-yl]pyridin-3-yl}pyrazine-2-carboxamide 356.8 0.83 190

3-amino-N-[4-(3-amino- pyrrolidin-1-yl)pyridin-3-yl]-pyrazine-2-carboxamide 300.1 191

3-amino-N-(4-piperazin-1-yl- pyridin-3-yl)pyrazine-2- carboxamide 300.1192

3-amino-N-{4-[(2R)-2-(amino- methyl)pyrrolidin-1-yl]pyridin-3-yl}pyrazine-2-carboxamide 314.1 193

3-amino-N-[4-(4-aminopiperidin- 1-yl)pyridin-3-yl]pyrazine-2-carboxamide 314.1 194

3-amino-N-[4-(2-amino- ethoxy)pyridin-3-yl]pyrazine-2- carboxamide 275.1195

3-amino-N-[2-(3-aminopiperidin- 1-yl)phenyl]pyrazine-2- carboxamide313.2 2.03 196

3-amino-N-[2-(3-aminopiperidin- 1-yl)pyridin-3-yl]pyrazine-2-carboxamide 314.2 1.66 197

N-[4-(3-aminopiperidin-1-yl)- pyridin-3-yl]pyrazine-2- carboxamide 299.20.86 198

N-[4-(3-aminopiperidin-1- yl)pyridin-3-yl]-5-methyl-pyrazine-2-carboxamide 313.2 1.05 199

3-amino-N-[4-(1,4-diazepan-1- yl)pyridin-3-yl]pyrazine-2- carboxamide314.1 0.55 200

2-amino-N-[4-(3-aminopiperidin- 1-yl)pyridin-3-yl]nicotinamide 313.2 201

3-amino-N-{2-[(3S)-3-amino- piperidin-1-yl]phenyl}pyrazine-2-carboxamide 313.12 2.39 202

3-amino-N-(2-piperazin-1- ylphenyl)pyrazine-2-carbox- amide 299.1 203

3-amino-N-[2-(4-aminopiperidin- 1-yl)phenyl]pyrazine-2- carboxamide313.2 204

3-amino-N-[2-(3-aminopiperidin- 1-yl)phenyl]-6-bromopyrazine-2-carboxamide 391 2.25 205

3-amino-N-[2-(4-aminopiperidin- 1-yl)phenyl]-6-bromopyrazine-2-carboxamide 391 2.2 206

3-amino-6-bromo-N-(2- piperazin-1-ylphenyl)pyrazine-2- carboxamide 3772.12 207

3-amino-N-[2-(4-aminopiperidin- 1-yl)-5-(1H-pyrazol-5-yl)phenyl]pyrazine-2-carbox- amide 379.1 2.01 208

3-amino-N-[2-(4-aminopiperidin- 1-yl)-5-fluorophenyl]pyrazine-2-carboxamide 331.1 2.47 209

3-amino-N-[2-(4-aminopiperidin- 1-yl)-6-fluorophenyl]pyrazine-2-carboxamide 331.2 1.99 210

3-amino-N-[2-(4-aminopiperidin- 1-yl)-4-fluorophenyl]pyrazine-2-carboxamide 331.1 2.38 211

3-amino-N-[2-(4-aminopiperidin- 1-yl)-5-methoxyphenyl]-6-bromopyrazine-2-carboxamide 421.1 2.74 212

3-amino-N-[2-(3-aminopiperidin- 1-yl)-5-methoxyphenyl]-6-bromopyrazine-2-carboxamide 421.1 2.6 213

3-amino-N-(5-chloro-2- piperazin-1-ylphenyl)pyrazine-2- carboxamide333.1 2.48 214

3-amino-N-[2-(4-aminopiperidin- 1-yl)-5-chlorophenyl]pyrazine-2-carboxamide 347.1 2.69 215

3-amino-N-[2-(3-aminopiperidin- 1-yl)-5-chlorophenyl]pyrazine-2-carboxamide 347.1 2.66 216

3-amino-N-(5-methyl-2- piperazin-1-ylphenyl)pyrazine-2- carboxamide313.2 2.36 217

3-amino-N-[2-(4-aminopiperidin- 1-yl)-5-methylphenyl]pyrazine-2-carboxamide 327.1 2.48 218

3-amino-N-[2-(3-aminopiperidin- 1-yl)-5-methylphenyl]pyrazine-2-carboxamide 327.1 2.55 219

N-(5-acetyl-2-piperazin-1-yl- phenyl)-3-aminopyrazine-2- carboxamide341.1 1.99 220

N-[5-acetyl-2-(4-aminopiperidin- 1-yl)phenyl]-3-aminopyrazine-2-carboxamide 355.2 2.11 221

N-[5-acetyl-2-(3-aminopiperidin- 1-yl)phenyl]-3-aminopyrazine-2-carboxamide 355.2 2.2 222

3-amino-N-[2-(4-aminopiperidin- 1-yl)-5-methoxyphenyl]pyrazine-2-carboxamide 343.1 2.33 223

3-amino-N-[2-(3-aminopiperidin- 1-yl)-5-methoxyphenyl]pyrazine-2-carboxamide 343.1 2.39 224

3-amino-N-[2-piperazin-1-yl-5- (trifluoromethyl)phenyl]pyrazine-2-carboxamide 367.1 2.72 225

3-amino-N-[2-(4-aminopiperidin- 1-yl)-5-(trifluoromethyl)phenyl]-pyrazine-2-carboxamide 381.1 2.87 226

3-amino-N-[2-(3-aminopiperidin- 1-yl)-5-(trifluoromethyl)phenyl]-pyrazine-2-carboxamide 381.1 2.88 227

3-amino-N-(4-methyl-2- piperazin-1-ylphenyl)pyrazine-2- carboxamide313.2 2.32 228

3-amino-N-[2-(4-aminopiperidin- 1-yl)-4-methylphenyl]pyrazine-2-carboxamide 327.2 2.46 229

3-amino-N-[2-(3-aminopiperidin- 1-yl)-4-methylphenyl]pyrazine-2-carboxamide 327.2 2.53 230

3-amino-N-[2-(3-aminopiperidin- 1-yl)-5-cyanophenyl]pyrazine-2-carboxamide 338.2 2.33 231

3-amino-pyrazine-2-carboxylic acid [2-(3-amino-piperidin-1-yl)-5-benzoyl-phenyl]-amide 417.1 2.79 232

3-amino-pyrazine-2-carboxylic acid (5-benzoyl-2-piperazin-1-yl-phenyl)-amide 403.2 2.7 233

3-amino-pyrazine-2-carboxylic acid [2-(4-amino-piperidin-1-yl)-5-benzoyl-phenyl]-amide 417.2 2.83 234

3-amino-N-[2-(4-aminopiperidin- 1-yl)-5-(methylsulfonyl)phenyl]-pyrazine-2-carboxamide 391.1 1.89 235

3-amino-N-[5-(3-aminopiperidin- 1-yl)-2-oxo-2,3-dihydro-pyrimidin-4-yl]pyrazine-2- carboxamide 331.1 1.18 236

3,5-diamino-N-{5-[3-(amino- methyl)piperidin-1-yl]-2-oxo-2,3-dihydropyrimidin-4-yl}-6- chloropyrazine-2-carboxamide 394.1 1.47237

3-amino-N-{5-[4-(3-amino- propanoyl)piperazin-1-yl]-2-oxo-2,3-dihydropyrimidin-4-yl}- pyrazine-2-carboxamide 388.1 1.18 238

3,5-diamino-6-chloro-pyrazine-2- carboxylic acid {5-[4-(3-amino-propionyl)-piperazin-1-yl]-2-oxo- 2,3-dihydro-pyrimidin-4-yl}- amide437.1 1.27 239

3-amino-N-[5-(3-aminopiperidin- 1-yl)-2-oxo-2,3-dihydro-pyrimidin-4-yl]-6-bromo- pyrazine-2-carboxamide 409 1.58 240

3-amino-N-{5-[3-(aminomethyl)- piperidin-1-yl]-2-oxo-2,3-dihydropyrimidin-4-yl}-6- bromopyrazine-2-carboxamide 423 1.72 241

3-amino-N-{5-[4-(3-amino- propanoyl)piperazin-1-yl]-2-oxo-2,3-dihydropyrimidin-4-yl}-6- bromopyrazine-2-carboxamide 466.1 1.49 242

3-amino-N-[5-(3-aminopiperidin- 1-yl)-2-oxo-2,3-dihydro-pyrimidin-4-yl]pyridine-2- carboxamide 330.1 1.28 243

3-amino-pyrazine-2-carboxylic acid [2-(3-amino-piperidin-1-yl)-5-cyclopropanecarbonyl-phenyl]- amide 381.1 2.49 244

3-amino-pyrazine-2-carboxylic acid [2-(4-amino-piperidin-1-yl)-5-cyclopropanecarbonyl-phenyl]- amide 381.1 2.45 245

3-amino-N-[5-(cyclopropyl- carbonyl)-2-piperazin-1-yl-phenyl]pyrazine-2-carboxamide 367.1 2.32 246

5-Amino-2-(2,6-difluoro- phenyl)-pyrimidine-4-carboxylic acid(3,5-diamino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)- amide441.2 1.30 247

5-Amino-2-(2-fluoro-phenyl)- pyrimidine-4-carboxylic acid(3,5-diamino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 423.21.39 248

3-Amino-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (3,5-diamino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 422.2 1.64249

N-{4-[(3S)-3-aminopiperidin- 1-yl]pyridin-3-yl}-2-(2,6-difluorophenyl)pyrimidine-4- carboxamide N-{4-[(3S)-3-aminopiperidin-1-yl]pyridin-3- yl}-2-(2,6-difluorophenyl)-pyrimidine-4-carboxamide 411.1 1.67 250

N-{4-[(3S)-3-aminopiperidin-1- yl]pyridin-3-yl}-2-(2-fluoro-phenyl)pyrimidine-4-carbox- amide 393.2 1.71 251

3-Amino-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid {4-[4-(3-amino-propionyl)-piperazin-1- yl]-pyridin-3-yl}-amide 464.2 1.98 252

3-Amino-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid {4-[4-(2-amino-acetyl)-piperazin-1-yl]- pyridin-3-yl}-amide 450.2 1.94 253

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid (3- amino-4-fluoro-3,4,5,6-tetra-hydro-2H-[1,4′]bipyridinyl-3′-yl)- amide 510.3 2.09 254

3-Amino-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (3-amino-4,4-difluoro-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 443.2 2.14 255

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid (3- amino-4,4-difluoro-3,4,5,6-tetra-hydro-2H-[1,4′]bipyridinyl-3′-yl)- amide 528.3 2.14 256

5-Amino-2-(2,6-difluoro- phenyl)-pyrimidine-4-carboxylic acid(3-amino-4-fluoro-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide444.1 1.71 257

3-amino-N-{4-[(3R,4R)-3-amino- 4-fluoropiperidin-1-yl]pyridin-3-yl}-6-(1,3-thiazol-2-yl)pyridine- 2-carboxamide 414.1 1.58 258

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid (3- amino-4-fluoro-3,4,5,6-tetra-hydro-2H-[1,4′]bipyridinyl-3′-yl)- amide 510.3 1.62 259

3-amino-N-{4-[(3S)-3-amino- piperidin-1-yl]pyridin-3-yl}-6-bromo-5-fluoropyridine-2- carboxamide 409.1 1.72 260

3-amino-N-{4-[(3S)-3-amino- piperidin-1-yl]pyridin-3-yl}-5-fluoropyridine-2-carboxamide 331.1 1.42 261

5-Amino-2-phenyl-pyrimidine-4- carboxylic acid (3-amino-3-methyl-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 404.2 1.90262

5-Amino-2-phenyl-pyrimidine-4- carboxylic acid (3-hydroxy-3-trifluoromethyl-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)- amide459.1 2.89 263

5-Amino-2-(2-fluoro-phenyl)- pyrimidine-4-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 408.2 1.78264

3-Amino-5-phenyl-pyridine-2- carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 389.3 2.12 265

5-Amino-2-phenyl-pyrimidine-4- carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 390.1 1.86 266

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid[4-(6-amino-2- trifluoromethyl-pyrimidin-4-yl)- pyridin-3-yl]-amide488.1 2.97 267

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid [4-(2- amino-6-methoxy-pyrimidin-4- yl)-pyridin-3-yl]-amide 517.22.69 268

5-Amino-2-phenyl-pyrimidine-4- carboxylic acid [4-(6-amino-2-methyl-pyrimidin-4-yl)-pyridin- 3-yl]-amide 399.2 2.13 269

5-Amino-2-phenyl-pyrimidine-4- carboxylic acid (6′-amino-2′-methyl-[4,4′]bipyridinyl-3-yl)- amide 398.2 2.02 270

5-Amino-2-(2,6-difluoro- phenyl)-pyrimidine-4-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 426.21.72 271

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid[4-(6-amino-2-methoxy- pyrimidin-4-yl)-pyridin-3-yl]- amide 450.2 2.70272

5-Amino-2-(2-fluoro-phenyl)- pyrimidine-4-carboxylic acid [4-(6-amino-2-methyl-pyrimidin-4- yl)-pyridin-3-yl]-amide 2.0 417.00 273

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid [4-(6- amino-2-methyl-pyrimidin-4-yl)- pyridin-3-yl]-amide 501.22.29 274

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid[4-(6-amino-2-methyl- pyrimidin-4-yl)-pyridin-3-yl]- amide 434.1 2.30275

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid[4-(6-amino-2- methylsulfanyl-pyrimidin-4-yl)- pyridin-3-yl]-amide 466.12.89 276

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid(3-amino-4-fluoro-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide443.2 2.05 277

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid(3-amino-4-fluoro-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide443.2 2.05 278

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid(5-amino-3-fluoromethyl- 3,4,5,6-tetrahydro-2H-[1,4′]bi-pyridinyl-3′-yl)-amide 457.1 2.17 279

3-Amino-6-cyclohexyl-pyridine- 2-carboxylic acid (3-amino-5-trifluoromethyl-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)- amide463.1 2.30 280

3-Fluoro-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (3-amino-5-trifluoromethyl-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 478.0 2.32 281

5-Fluoro-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (3-amino-5-trifluoromethyl-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 478.1 2.42 282

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid(3-amino-5-trifluoromethyl- 3,4,5,6-tetrahydro-2H-[1,4′]bi-pyridinyl-3′-yl)-amide 493.1 2.36 283

3-Amino-6-cyclohexyl-pyridine- 2-carboxylic acid (3-amino-5-methyl-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 409.2 2.44284

3-Amino-6-cyclohexyl-pyridine- 2-carboxylic acid (3-amino-5-fluoro-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 413.1 2.27285

3-Amino-6-cyclohexyl-pyridine- 2-carboxylic acid (3-amino-5-methoxy-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 425.2 2.31286

3-Fluoro-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (5-amino-3-methyl-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)- amide424.1 2.20 287

5-Fluoro-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (3-amino-5-methyl-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)- amide424.1 2.31 288

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid(3-amino-5-methyl-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide439.1 2.27 289

5-Fluoro-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (5-amino-3-hydroxymethyl-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 440.1 1.94 290

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid[4-(2,7-diaza-spiro[4.5]dec- 7-yl)-pyridin-3-yl]-amide 465.1 2.12 291

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid [4- (2,7-diaza-spiro[4.5]dec-7-yl)- pyridin-3-yl]-amide 532.1 2.02292

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid (3- amino-5-fluoro-3,4,5,6-tetra-hydro-2H-[1,4′]bipyridinyl-3′-yl)- amide 510.0 2.12 293

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid(3-amino-5-fluoro-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide443.0 2.07 294

Benzoic acid 5-amino-3′-{[3- amino-6-(2-fluoro-5-isopropylcarbamoyl-phenyl)- pyridine-2-carbonyl]-amino}-3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3-yl ester 612.1 2.49 295

Benzoic acid 5-amino-3′-{[3- amino-6-(2,6-difluoro-phenyl)-pyridine-2-carbonyl]-amino}- 3,4,5,6-tetrahydro-2H-[1,4′]bi-pyridinyl-3-yl ester 545.0 2.51 296

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid (3- amino-5-hydroxy-3,4,5,6-tetra-hydro-2H-[1,4′]bipyridinyl-3′-yl)- amide 508.2 2.00 297

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid(3-amino-5-hydroxy-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 441.1 1.80 298

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid (3- amino-5-methoxy-3,4,5,6-tetra-hydro-2H-[1,4′]bipyridinyl-3′-yl)- amide 522.2 2.01 299

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid (3- amino-4-methoxy-3,4,5,6-tetra-hydro-2H-[1,4′]bipyridinyl-3′-yl)- amide 522.0 2.09 300

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid(3-amino-5-methoxy- 3,4,5,6-tetrahydro-2H-[1,4′]bi-pyridinyl-3′-yl)-amide 455.2 1.98 301

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid(3-amino-4-methoxy- 3,4,5,6-tetrahydro-2H-[1,4′]bi-pyridinyl-3′-yl)-amide 455.0 1.96 302

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid[4-(2-oxo-hexahydro- oxazolo[4,5-c]pyridin-5-yl)- pyridin-3-yl]-amide467.0 2.40 303

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid[4-(2-oxo-hexahydro- oxazolo[4,5-c]pyridin-5-yl)- pyridin-3-yl]-amide467.0 2.40

Example 304 Synthesis ofN-(4-(3-acetamidopiperidin-1-yl)pyridin-3-yl)-3-amino-6-bromopyrazine-2-carboxamide

To solution of3-amino-N-(4-(3-aminopiperidin-1-yl)pyridin-3-yl)-6-bromopyrazine-2-carboxamidein CH₂Cl₂ at a concentration of 0.5 M at room temperature was addedtriethylamine (3 eq) followed by acetic anhydride (1.2 eq). The reactionwas stirred at room temperature for 30 minutes, concentrated, purifiedby reverse phase HPLC and lyophilized to provideN-(4-(3-acetamidopiperidin-1-yl)pyridin-3-yl)-3-amino-6-bromopyrazine-2-carboxamide,as the TFA salt. LCMS (m/z): 434.1 (MH⁺).

Method 11 Example 305 Synthesis of tert-butyl1-(3-(3-amino-6-bromopyrazine-2-carboxamido)pyridin-4-yl)piperidin-3-ylcarbamate

A solution containing 1 eq each of tert-butyl1-(3-aminopyridin-4-yl)piperidin-3-ylcarbamate,3-amino-6-bromopyrazine-2-carboxylic acid, HOAT and EDC in DMF, at aconcentration of 0.5 M, was stirred for 60 hours. The solution wasdiluted with EtOAc and was washed with H₂O (4×), NaCl_((sat)), was driedover MgSO₄, was filtered and the volatiles were removed in vacuo. Afterpurification by silica gel chromatography (EtOAc eluant), tert-butyl1-(3-(3-amino-6-bromopyrazine-2-carboxamido)pyridin-4-yl)piperidin-3-ylcarbamatewas obtained (78%). LCMS (m/z): 492.2 (MH⁺); LC R_(t)=2.68 min.

Example 306 Synthesis of tert-butyl14243-amino-6-bromopyrazine-2-carboxamido)-4-benzoylphenyl)piperidin-4-ylcarbamate

Following method 11, tert-butyl1-(2-amino-4-benzoylphenyl)piperidin-4-ylcarbamate was coupled to3-amino-6-bromopyrazine-2-carboxylic acid with TEA (1.5 eq) in ACN at55° C. for 48 hours. Concentrated, triturated in cold ACN, filtered, anddried in vacuo yielding tert-butyl1-(2-(3-amino-6-bromopyrazine-2-carboxamido)-4-benzoylphenyl)piperidin-4-ylcarbamate(46%). LCMS (m/z): 595.2 (MH⁺); LC R_(t)=3.94 min.

Example 307 Synthesis of tert-butyl1-(2-(3-amino-6-bromopyrazine-2-carboxamido)-4-benzoylphenyl)piperidin-3-ylcarbamate

Following method 11, tert-butyl1-(2-amino-4-benzoylphenyl)piperidin-3-ylcarbamate was coupled to3-amino-6-bromopyrazine-2-carboxylic acid with TEA (1.5 eq) in ACN at55° C. for 48 hours. Concentrated, triturated in cold ACN, filtered, anddried in vacuo yielding tert-butyl1-(2-(3-amino-6-bromopyrazine-2-carboxamido)-4-benzoylphenyl)piperidin-3-ylcarbamate(30%). LCMS (m/z): 595.1 (MH⁺); LC R_(t)=3.87 min.

Example 308 Synthesis of tert-butyl4-(2-(3-amino-6-bromopyrazine-2-carboxamido)-4-benzoylphenyl)piperazine-1-carboxylate

Following method 11, tert-butyl4-(2-amino-4-benzoylphenyl)piperazine-1-carboxylate was coupled to3-amino-6-bromopyrazine-2-carboxylic acid with TEA (1.5 eq) in ACN at55° C. for 48 hours. Concentrated, triturated in cold ACN, filtered, anddried in vacuo yielding tert-butyl4-(2-(3-amino-6-bromopyrazine-2-carboxamido)-4-benzoylphenyl)piperazine-1-carboxylate(50%). LCMS (m/z): 581.1 (MH⁺); LC R_(t)=4.00 min.

Example 309 Synthesis of tert-butyl1-(2-(3-amino-6-bromopyrazine-2-carboxamido)-4-methoxyphenyl)piperidin-4-ylcarbamate

Following method 11, tert-butyl1-(2-amino-4-methoxyphenyl)piperidin-4-ylcarbamate was coupled to3-amino-6-bromopyrazine-2-carboxylic acid with TEA (3 eq) in ACN at 55°C. for 48 hours. Concentrated, triturated in cold ACN, filtered, anddried in vacuo yielding tert-butyl1-(2-(3-amino-6-bromopyrazine-2-carboxamido)-4-methoxyphenyl)piperidin-4-ylcarbamate(7%). LCMS (m/z): 521.1 (MH⁺); LC R_(t)=3.63 min.

Example 310 Synthesis of 3,5-diamino-6-chloropyrazine-2-carboxylic acid

To a solution of methyl 3,5-diamino-6-chloropyrazine-2-carbamate (5 g,0.025 mols) in 2:1 THF/MeOH (90 mL) was added 1M LiOH (62 mL, 0.062mols). After the reaction was stirred at r.t. 72 hrs, 1N HCl (62 mL,0.062 mols) was added. The reaction was filtered and washed with water(3×10 mL) to give 3,5-diamino-6-chloropyrazine-2-carboxylic acid aswhite solid 4.3 g (93% yield). LCMS (m/z): 189.1 (MH⁺); LC R_(t)=1.05min.

Example 311 Synthesis of3,5-diamino-6-chloro-N-(4-(piperidin-1-yl)pyridin-3-yl)pyrazine-2-carboxamide

Following method 11, 4-(piperidin-1-yl)pyridin-3-amine was coupled to3,5-diamino-6-chloropyrazine-2-carboxylic acid yielding3,5-diamino-6-chloro-N-(4-(piperidin-1-yl)pyridin-3-yl)pyrazine-2-carboxamide(76%). LCMS (m/z): 347.8 (MH⁺); LC R_(t)=2.17 min.

Example 312 Synthesis of3,5-diamino-N-(4-(piperidin-1-yl)pyridin-3-yl)pyrazine-2-carboxamide

Following method 2 where diethylamine (4.0 eq) was also included and thereaction was recharged with Pd/C and H₂ after 2 and 4 days,3,5-diamino-6-chloro-N-(4-(piperidin-1-yl)pyridin-3-yl)pyrazine-2-carboxamidewas reduced in 7 days yielding3,5-diamino-N-(4-(piperidin-1-yl)pyridin-3-yl)pyrazine-2-carboxamide asthe TFA salt. LCMS (m/z): 314.1 (MH⁺); LC R_(t)=1.67 min.

Example 313 Synthesis of tert-butyl1-(3-(3,5-diamino-6-chloropyrazine-2-carboxamido)pyridin-4-yl)piperidin-3-ylcarbamate

Following method 11, tert-butyl1-(3-aminopyridin-4-yl)piperidin-3-ylcarbamate was coupled to3,5-diamino-6-chloropyrazine-2-carboxylic acid yielding (S)-tert-butyl1-(3-(3,5-diamino-6-chloropyrazine-2-carboxamido)pyridin-4-yl)piperidin-3-ylcarbamate(57%). LCMS (m/z): 463.1 (MH⁺); LC R_(t)=2.36 min.

Example 314 Synthesis of 3-amino-6-bromopicolinic acid

To a solution of methyl 3-amino-6-bromopicolinate (2.31 g, 10 mmoles) in2:1 THF/MeOH (51 mL) was added 1.0 M LiOH (17 mL, 17 mmoles). Afterstirring for 16 hours, 1 N HCl (17 mL, 17 mmoles) was added and theTHF/MeOH was removed in vacuo. The resulting solid was filtered, rinsedwith cold H₂O (4×20 mL) and pumped on yielding 3-amino-6-bromopicolinicacid (97%). LCMS (m/z): 216.9 (MH⁺); LC R_(t)=1.93 min.

Example 315 Synthesis of (S)-tert-butyl1-(3-(3-amino-6-bromopicolinamido)pyridin-4-yl)piperidin-3-ylcarbamate

Following method 11, (S)-tert-butyl1-(3-aminopyridin-4-yl)piperidin-3-ylcarbamate was coupled to3-amino-6-bromopicolinic acid and purified by SiO₂ chromatography (EtOAceluant) yielding (S)-tert-butyl1-(3-(3-amino-6-bromopicolinamido)pyridin-4-yl)piperidin-3-ylcarbamate(45%). LCMS (m/z): 491.1 (MH⁺); LC R_(t)=2.89 min.

Method 12 Example 316 Synthesis of3-amino-N-(4-(3-aminopiperidin-1-yl)pyridin-3-yl)-6-(2-chlorophenyl)pyrazine-2-carboxamide

A solution of tert-butyl1-(3-(3-amino-6-bromopyrazine-2-carboxamido)pyridin-4-yl)piperidin-3-ylcarbamate(1.0 eq), ortho-chlorophenyl boronic acid (3.0 eq.), Pd(dppf)Cl₂—CH₂Cl₂and triethylamine (9.0 eq.) in dimethylacetamide (concentration=0.1 M)was heated at 130° C. with microwave irradiation for 900 seconds. Uponcooling the N-Boc Suzuki product was directly purified by reverse phaseHPLC. The product fraction was lyophilized and the resulting solid wastreated with 25% TFA/DCM (at a resulting concentration of 0.05 M). Aftersitting for 2 hours, the volatiles were removed in vacuo and the residuewas purified by reverse phase HPLC. After lyophilization,3-amino-N-(4-(3-aminopiperidin-1-yl)pyridin-3-yl)-6-(2-chlorophenyl)pyrazine-2-carboxamidewas obtained (56%) as the TFA salt. LCMS (m/z): 424.1 (MH⁺); LCR_(t)=1.94 min.

Alternatively, the free base and HCl salt could be obtained as describedin Method 9.

The following compounds were prepared using Method 12. In some instancesNMP or DMF was used in place of dimethylacetamide.

Example Structure Name MH+ LC 317

3-amino-6-(2-fluoro-5-iso- propylcarbamoyl-phenyl)-pyridine-2-carboxylic acid ((S)-3-amino-3,4,5,6-tetra-hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 492.2 2.04 318

3-amino-6-[2-methyl-5- (piperidine-1-sulfonyl)-phenyl]-pyridine-2-carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 550.3 2.44 319

3-amino-6-(4-methane- sulfonyl-phenyl)-pyridine-2- carboxylic acid((S)-3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide467.2 1.77 320

3-amino-6-(3-methane- sulfonyl-phenyl)-pyridine-2- carboxylic acid((S)-3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3-yl)-amide467.2 1.77 321

3-amino-6-[2-methyl-5- (pyrrolidine-1-sulfonyl)-phenyl]-pyridine-2-carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 536.2 2.27 322

3-amino-6-(2-chloro-5- ethoxy-phenyl)-pyridine-2- carboxylic acid((S)-3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide467.2 2.43 323

3-amino-6-(2-fluoro-5- phenylcarbamoyl-phenyl)- pyridine-2-carboxylicacid ((S)-3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 526.2 2.35 324

3-amino-6-(5-cyclohexyl- carbamoyl-2-fluoro-phenyl)-pyridine-2-carboxylic acid ((S)-3-amino-3,4,5,6-tetra-hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 532.3 2.44 325

4-[5-amino-6-((S)-3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi-pyridinyl-3′-ylcarbamoyl)- pyridin-2-yl]-3-fluoro-benzoic acid methylester 465.2 2.24 326

3-amino-6-(2-fluoro-5-propyl- carbamoyl-phenyl)-pyridine- 2-carboxylicacid ((S)-3- amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide492.2 2.11 327

3-amino-6-(5-ethylcarbamoyl- 2-fluoro-phenyl)-pyridine-2- carboxylicacid ((S)-3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi-pyridinyl-3′-yl)-amide 478.2 1.97 328

3-amino-6-(5-dimethyl- carbamoyl-2-fluoro-phenyl)- pyridine-2-carboxylicacid ((S)-3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 478.2 1.98 329

3-amino-6-(5-diethyl- sulfamoyl-2-methyl-phenyl)- pyridine-2-carboxylicacid ((S)-3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 538.3 2.41 330

3-amino-6-(5-dimethyl- sulfamoyl-2-methyl-phenyl)- pyridine-2-carboxylicacid ((S)-3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 510.2 2.12 331

5-amino-2′-chloro-[2,3′]bi- pyridinyl-6-carboxylic acid((S)-3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide424.1 1.78 332

3-amino-6-(2-cyano-phenyl)- pyridine-2-carboxylic acid((S)-3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide414.2 1.87 333

3-amino-6-(2-fluoro-5- methyl-phenyl)-pyridine-2- carboxylic acid((S)-3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide421.2 2.29 334

3-amino-6-(2-fluoro-5- methylcarbamoyl-phenyl)- pyridine-2-carboxylicacid ((S)-3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 464.2 1.86 335

3-amino-6-(5-tert-butyl- sulfamoyl-2-methyl-phenyl)-pyridine-2-carboxylic acid ((S)-3-amino-3,4,5,6-tetra-hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 538.3 2.36 336

3-amino-6-(5-tert-butyl- sulfamoyl-2-methyl-phenyl)-pyridine-2-carboxylic acid ((S)-3-amino-3,4,5,6-tetra-hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 457.2 2.28 337

3-amino-6-(2-methoxy- phenyl)-pyridine-2-carboxylic acid((S)-3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bi- pyridinyl-3-yl)-amide419.2 2.11 338

3-amino-6-o-tolyl-pyridine-2- carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 403.2 2.17 339

5-amino-3′-fluoro-[2,4′]bi- pyridinyl-6-carboxylic acid((S)-3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide408.2 1.48 340

3,5-diamino-6-o-tolyl- pyrazine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 419.2 1.67341

3,5-diamino-6-(2-trifluoro- methyl-phenyl)-pyrazine-2- carboxylic acid(3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 473.21.76 342

3,5-diamino-6-(2-chloro- phenyl)-pyrazine-2-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 439.11.68 343

3,5-diamino-6-(2-fluoro- phenyl)-pyrazine-2-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 423.11.59 344

3,5-diamino-6-(2-methoxy- phenyl)-pyrazine-2-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 435.21.58 345

3-amino-6-(2-cyano-phenyl)- pyrazine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 415.2 1.67346

3-amino-6-(2-chloro-phenyl)- pyrazine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 424.1 1.94347

3-amino-6-(2-trifluoro- methoxy-phenyl)-pyrazine-2- carboxylic acid(3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 474.22.16 348

3-amino-6-(2-trifluoromethyl- phenyl)-pyrazine-2-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 458.22.03 349

3-amino-6-(2-methoxy- phenyl)-pyrazine-2-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 420.21.86 350

3-amino-6-(2-fluoro-phenyl)- pyrazine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 408.1 1.84351

3-amino-6-o-tolyl-pyrazine-2- carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 404.2 1.9 352

3-amino-6-(3-chloro-phenyl)- pyrazine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 424.1 353

3-amino-6-(4-chloro-phenyl)- pyrazine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 424.1 354

3-amino-6-pyridin-4-yl- pyrazine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 391.2 355

3-amino-6-(1H-pyrazol-4-yl)- pyrazine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 380.1 356

3-amino-6-pyridin-3-yl- pyrazine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 391.2 357

3-amino-6-pyrimidin-5-yl- pyrazine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 392.2 358

3-amino-6-(4-hydroxy- phenyl)-pyrazine-2-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 406.21.58 359

3-amino-6-phenyl-pyrazine-2- carboxylic acid (4-piperazin-1-yl-pyridin-3-yl)-amide 376.2 1.68 360

5-amino-6′-methoxy- [2,2′]bipyrazinyl-6-carboxylic acid(4-piperazin-1-yl- pyridin-3-yl)-amide 408.2 1.6 361

3-amino-6-(3-carbamoyl- phenyl)-pyrazine-2-carboxylic acid(4-piperazin-1-yl- pyridin-3-yl)-amide 419.2 1.36 362

3-amino-6-(4-methoxy- phenyl)-pyrazine-2-carboxylic acid(4-piperazin-1-yl- pyridin-3-yl)-amide 406.2 1.76 363

3-amino-6-(2-fluoro-phenyl)- pyrazine-2-carboxylic acid (4-piperazin-1-yl-pyridin-3-yl)- amide 394.2 1.74 364

3-amino-6-phenyl-pyrazine-2- carboxylic acid (4-amino-3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 390.2 1.73 365

3-amino-6-(2-phenoxy- phenyl)-pyrazine-2-carboxylic acid(4-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 482.22.33 366

5-amino-6′-methoxy-[2,2′]bi- pyrazinyl-6-carboxylic acid(4-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)- amide 422.21.64 367

3-amino-6-(4-methoxy- phenyl)-pyrazine-2-carboxylic acid(4-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 420.21.8 368

3-amino-6-(2-fluoro-phenyl)- pyrazine-2-carboxylic acid (4-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 408.2 1.79369

3-amino-6-(3-carbamoyl- phenyl)-pyrazine-2-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 433.21.51 370

3-amino-6-(4-methoxy- phenyl)-pyrazine-2-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 420.21.8 371

3-amino-6-methyl-pyrazine-2- carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 328.2 1.23 372

3-amino-6-phenyl-pyrazine-2- carboxylic acid [2-(3-amino-piperidin-1-yl)-phenyl]-amide 389.2 2.99 373

3-amino-6-(3-carbamoyl- phenyl)-pyrazine-2-carboxylic acid[2-(3-amino-piperidin-1- yl)-phenyl]-amide 432.1 2.54 374

3-amino-6-(2-fluoro-phenyl)- pyrazine-2-carboxylic acid [2-(3-amino-piperidin-1-yl)- phenyl]-amide 407.2 3.03 375

3-amino-6-(4-methoxy- phenyl)-pyrazine-2-carboxylic acid[2-(3-amino-piperidin-1- yl)-phenyl]-amide 419.2 3.06 376

3-amino-6-methyl-pyrazine-2- carboxylic acid [2-(3-amino-piperidin-1-yl)-phenyl]-amide 327.2 2.44 377

3-amino-6-(2-fluoro-phenyl)- pyrazine-2-carboxylic acid [2-(4-amino-piperidin-1-yl)- phenyl]-amide 407.2 2.94 378

3-amino-N-[2-(4-amino- piperidin-1-yl)phenyl]-6- methylpyrazine-2-carboxamide 327.2 2.36 379

3-amino-6-furan-3-yl- pyrazine-2-carboxylic acid [2-(4-amino-piperidin-1-yl)- phenyl]-amide 379.2 2.64 380

3-amino-6-(3-carbamoyl- phenyl)-pyrazine-2-carboxylic acid[2-(4-amino-piperidin-1- yl)-phenyl]-amide 432.2 2.56 381

3-amino-6-(4-methoxy- phenyl)-pyrazine-2-carboxylic acid[2-(4-amino-piperidin-1- yl)-phenyl]-amide 419.1 2.96 382

3-amino-6-phenyl-pyrazine-2- carboxylic acid (2-piperazin-1-yl-phenyl)-amide 375.1 2.76 383

3-amino-6-(2-fluoro-phenyl)- pyrazine-2-carboxylic acid (2-piperazin-1-yl-phenyl)-amide 393.1 2.8 384

3-amino-6-furan-3-yl- pyrazine-2-carboxylic acid (2-piperazin-1-yl-phenyl)-amide 365.1 2.71 385

3-amino-6-(3-carbamoyl- phenyl)-pyrazine-2-carboxylic acid(2-piperazin-1-yl- phenyl)-amide 418.2 2.36 386

3-amino-6-(4-methoxy- phenyl)-pyrazine-2-carboxylic acid(2-piperazin-1-yl- phenyl)-amide 405.1 3 387

3-amino-6-(2-phenoxy- phenyl)-pyrazine-2-carboxylic acid(2-piperazin-1-yl- phenyl)-amide 467.2 3.28 388

3-amino-6-m-tolyl-pyrazine- 2-carboxylic acid (2-piper-azin-1-yl-phenyl)-amide 389.2 2.95 389

3-amino-6-naphthalen-1-yl- pyrazine-2-carboxylic acid (2-piperazin-1-yl-phenyl)-amide 425.2 3.06 390

3-amino-6-(2-methoxy- pyridin-3-yl)-pyrazine-2- carboxylic acid(2-piperazin- 1-yl-phenyl)-amide 406.2 2.8 391

3-amino-6-(2-methoxy- pyrimidin-5-yl)-pyrazine-2- carboxylic acid(2-piperazin- 1-yl-phenyl)-amide 407.2 2.55 392

3-amino-6-[3-(morpholine-4- carbonyl)-phenyl]-pyrazine-2- carboxylicacid [2-(4-amino- piperidin-1-yl)-phenyl]-amide 502.1 2.71 393

3-amino-6-(2-fluoro-5- methoxy-phenyl)-pyrazine-2- carboxylic acid[2-(4-amino- piperidin-1-yl)-phenyl]-amide 437.1 3.05 394

3-amino-6-(3,4,5-trimethoxy- phenyl)-pyrazine-2-carboxylic acid[2-(4-amino-piperidin-1- yl)-phenyl]-amide 479.2 2.89 395

3-amino-6-(3,4,5-trimethoxy- phenyl)-pyrazine-2-carboxylic acid(2-piperazin-1-yl- phenyl)-amide 465.2 2.74 396

3-amino-6-(2-fluoro-5- methoxy-phenyl)-pyrazine-2- carboxylic acid(2-piperazin- 1-yl-phenyl)-amide 423.2 2.96 397

3-amino-6-phenyl-pyrazine-2- carboxylic acid [2-(4-amino-piperidin-1-yl)-phenyl]-amide 389.2 2.37 398

3-amino-6-[3-(morpholine-4- carbonyl)-phenyl]-pyrazine-2- carboxylicacid (2-piperazin- 1-yl-phenyl)-amide 488.2 2.5 399

3-amino-6-phenyl-pyrazine-2- carboxylic acid [2-(4-amino-piperidin-1-yl)-phenyl]-amide 389.2 2.37 400

3-amino-6-o-tolyl-pyrazine-2- carboxylic acid [5-(3-amino-piperidin-1-yl)-2-oxo-2,3- dihydro-pyrimidin-4-yl]-amide 421.1 1.96 401

3-amino-6-(2-fluoro-phenyl)- pyrazine-2-carboxylic acid [5-(3-amino-piperidin-1-yl)-2- oxo-2,3-dihydro-pyrimidin-4- yl]-amide 425.11.92 402

3-amino-6-(2-chloro-phenyl)- pyrazine-2-carboxylic acid [5-(3-amino-piperidin-1-yl)-2- oxo-2,3-dihydro-pyrimidin-4- yl]-amide 441.12 403

3-amino-6-(2-cyano-phenyl)- pyrazine-2-carboxylic acid [5-(3-amino-piperidin-1-yl)-2- oxo-2,3-dihydro-pyrimidin-4- yl]-amide 432.11.7 404

3-amino-6-(2-trifluoromethyl- phenyl)-pyrazine-2-carboxylic acid[5-(3-amino-piperidin-1- yl)-2-oxo-2,3-dihydro- pyrimidin-4-yl]-amide475.1 2.09 405

3-amino-6-(2-trifluoro- methoxy-phenyl)-pyrazine-2- carboxylic acid[5-(3-amino- piperidin-1-yl)-2-oxo-2,3- dihydro-pyrimidin-4-yl]-amide491.1 2.19

Method 13 Example 406 Synthesis of(S)-3-amino-N-(4-(3-aminopiperidin-1-yl)pyridin-3-yl)-6-(2-fluoro-4-methylphenyl)picolinamide

A solution of (S)-tert-butyl1-(3-(3-amino-6-bromopicolinamido)pyridin-4-yl)piperidin-3-ylcarbamate(1.0 eq), 2-fluoro-4-methyl phenyl boronic acid (3.0 eq.),Pd(dppf)Cl₂—CH₂Cl₂ (0.15 eq.) in 3:1 DME/2M Na₂CO₃ (concentration=0.1 M)was heated at 120° C. with microwave irradiation for 1200 seconds. Uponcooling the organic layer was separated, concentrated and the N-BocSuzuki product was directly purified by reverse phase HPLC. The productfraction was lyophilized and the resulting solid was treated with 25%TFA/DCM (at a resulting concentration of 0.05 M). After sitting for 2hours, the volatiles were removed in vacuo and the residue was purifiedby reverse phase HPLC. After lyophilization,(S)-3-amino-N-(4-(3-aminopiperidin-1-yl)pyridin-3-yl)-6-(2-fluoro-4-methylphenyl)picolinamidewas obtained (44%) as the TFA salt. LCMS (m/z): 2.23 (MH⁺); LCR_(t)=421.2 min.

Alternatively, the free base and HCl salt of(S)-3-amino-N-(4-(3-aminopiperidin-1-yl)pyridin-3-yl)-6-(2-fluoro-4-methylphenyl)picolinamidecould be obtained as described in Method 9.

The following compounds were prepared using Method 13.

Example Structure Name MH+ LC 407

3-amino-6-pyrimidin-5-yl- pyridine-2-carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 391.2 1.4 408

5,2′-diamino-6′-fluoro-[2,3′]bi- pyridinyl-6-carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 423.21.79  409

3-amino-6-(2-amino-4-methyl- pyrimidin-5-yl)-pyridine-2- carboxylic acid((S)-3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide420.2 1.21  410

5,6′-diamino-4′-chloro-[2,3′]bi- pyridinyl-6-carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 439.21.41  411

3-amino-6-(2-amino-pyrimidin-5- yl)-pyridine-2-carboxylic acid((S)-3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 406.21.25  412

5,2′-diamino-[2,4′]bipyridinyl-6- carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 405.2 1.26  413

5-amino-6′-methoxy-[2,2′]bi- pyridinyl-6-carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 420.21.87  414

5-amino-[2,3′]bipyridinyl-6- carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 390.2 1.11  415

5-amino-[2,4′]bipyridinyl-6- carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 390.2 1.11  416

5,6′-diamino-5′-methoxy-[2,3′]bi- pyridinyl-6-carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 435.21.34  417

3-amino-6-(2,4-diamino- pyrimidin-5-yl)-pyridine-2- carboxylic acid((S)-3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide421.2 1.07  418

5,6′-diamino-[2,3′]bipyridinyl-6- carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 405.2 1.26  419

5-amino-2′-fluoro-[2,3′]bi- pyridinyl-6-carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 408.2 1.7 420

3-amino-6-(4-chloro-2-fluoro- phenyl)-pyridine-2-carboxylic acid((S)-3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)- amide441.2 2.29  421

5-amino-4′-methoxy-[2,3′]bi- pyridinyl-6-carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 420.2 1.7 422

5-amino-3′-chloro-[2,4′]bi- pyridinyl-6-carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 424.11.61  423

5-amino-2′-methoxy-[2,3′]bi- pyridinyl-6-carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 420.21.81  424

3-amino-6-(2,3-difluoro-phenyl)- pyridine-2-carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 425.2 2.15 425

3-amino-6-(2,6-difluoro-phenyl)- pyridine-2-carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 425.2 2.03 426

3-amino-6-(2,6-dimethyl- phenyl)-pyridine-2-carboxylic acid((S)-3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)- amideQ417.2 2.2  427

3-amino-6-(5-fluoro-2-methoxy- phenyl)-pyridine-2-carboxylic acid((S)-3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)- amide437.2 2.11  428

3-amino-6-(4-fluoro-2-methoxy- phenyl)-pyridine-2-carboxylic acid((S)-3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)- amide437.2 2.15  429

3-amino-6-(4-chloro-2-methyl- phenyl)-pyridine-2-carboxylic acid((S)-3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)- amide437.2 2.39  430

3-amino-6-(4-fluoro-2-methyl- phenyl)-pyridine-2-carboxylic acid((S)-3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)- amide421.2 2.23  431

3-amino-6-(2,4-difluoro-phenyl)- pyridine-2-carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 425.2 2.15 432

3-amino-6-(2-fluoro-4-methyl- phenyl)-pyridine-2-carboxylic acid((S)-3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)- amide421.2 2.27  433

3-amino-6-(2-chloro-phenyl)- pyridine-2-carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 423.2 2.15 434

3-amino-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid ((S)-3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 407.2 2.07 435

3-amino-6-phenyl-pyrazine-2- carboxylic acid [2-(3-amino-piperidin-1-yl)-5-benzoyl- phenyl]-amide 493.2 3.35  436

3-amino-6-phenyl-pyrazine-2- carboxylic acid [2-(4-amino-piperidin-1-yl)-5-benzoyl- phenyl]-amide 493.2 3.3  437

3-amino-6-phenyl-pyrazine-2- carboxylic acid [2-(4-amino-piperidin-1-yl)-5-methoxy- phenyl]-amide 419.1 3.07  438

3-amino-6-(2-fluoro-phenyl)- pyrazine-2-carboxylic acid (5-benzoyl-2-piperazin-1-yl- phenyl)-amide 497.2 3.25  439

3-amino-6-(2-fluoro-phenyl)- pyrazine-2-carboxylic acid [2-(4-amino-piperidin-1-yl)-5-benzoyl- phenyl]-amide 511.1 3.29  440

3-amino-6-methyl-pyrazine-2- carboxylic acid (5-benzoyl-2-piperazin-1-yl-phenyl)-amide 417.2 2.75  441

3-amino-6-(4-methoxy-phenyl)- pyrazine-2-carboxylic acid (5-benzoyl-2-piperazin-1-yl- phenyl)-amide 509.3 3.28  442

5-Fluoro-6-phenyl-pyridine-2- carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 392.2 2.12  443

5-Fluoro-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)-pyridine-2-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 495.2 2.13  444

5-Fluoro-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 410.1 2.15 445

6-(2,6-Difluoro-phenyl)-5-fluoro- pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3-yl)-amide 428.1 2.14 446

3-Amino-6-[2,2′]bithiophenyl-5- yl-pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 477.1 2.54 447

3-Amino-6-(2,3-dihydro- thieno[3,4-b][1,4]dioxin-5-yl)-pyridine-2-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 453.1 2.02  448

3-Amino-6-(3-methyl-thiophen- 2-yl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 409.22.10  449

3-Amino-6-thiophen-3-yl- pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 395.1 1.95 450

3-Amino-6-(5-chloro-thiophen-2- yl)-pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 429.1 2.23 451

3-Amino-6-(4-methyl-thiophen- 2-yl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 409.22.14  452

3-Amino-6-(3-ethyl-phenyl)- pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 417.2 2.43 453

6-(2-Fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylic acid(3- amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 477.22.06  454

6-(2,6-Difluoro-phenyl)-pyridine- 2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 410.1 2.02  455

6-(2-Fluoro-phenyl)-pyridine-2- carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 392.2 2.10  456

3-Amino-6-(3-isopropyl-phenyl)- pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 431.2 2.61 457

3-Amino-6-(3-cyanomethyl- phenyl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 428.22.06  458

3-Amino-6-biphenyl-3-yl- pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 465.2 2.71 459

3-Amino-6-(3-bromo-phenyl)- pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 467.1 2.30 460

3-Amino-6-(3-trifluoromethyl- phenyl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 457.22.41  461

3-Amino-6-(3-cyano-phenyl)- pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 414.2 2.02 462

3-Amino-6-(2,6-difluoro-phenyl)- pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 425.2 2.26 463

3-Amino-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 407.2 2.29 464

3-Amino-6-[3-(2-chloro- benzyloxy)-2,6-difluoro-phenyl]-pyridine-2-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 565.2 2.98  465

3-Amino-6-[2,6-difluoro-3-(2- fluoro-benzyloxy)-phenyl]-pyridine-2-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 549.2 2.70  466

3-Amino-6-(3-butoxy-2,6- difluoro-phenyl)-pyridine-2- carboxylic acid(3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 497.32.83  467

3-Amino-6-(2-fluoro-5- isopropoxy-phenyl)-pyridine-2- carboxylic acid(3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 465.32.59  468

3-Amino-6-(2-fluoro-5-propoxy- phenyl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 465.22.68  469

3-Amino-6-(5-ethoxy-2-fluoro- phenyl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 451.22.47  470

3-Amino-6-(2-fluoro-5-methoxy- phenyl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 437.22.33  471

3-Amino-6-(2-fluoro-3-methoxy- phenyl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 437.22.26  472

5-Amino-4′-methyl-[2,3′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 404.2 1.18 473

3-Amino-6-biphenyl-4-yl- pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 465.2 2.71 474

3-Amino-6-(3-chloro-thiophen-2- yl)-pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 429.1 2.21 475

3-Amino-6-thiophen-2-yl- pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 395.2 1.97 476

5-Amino-2′-fluoro-6′-methyl- [2,3′]bipyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 422.21.93  477

3-Amino-6-(2,5-difluoro-4- methoxy-phenyl)-pyridine-2- carboxylic acid(3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 455.22.29  478

3-Amino-6-(5-chloro-2-fluoro-4- methyl-phenyl)-pyridine-2- carboxylicacid (3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide455.2 2.48  479

3-Amino-6-(3-chloro-2-fluoro- phenyl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 441.22.36  480

3-Amino-6-(4-sulfamoyl- phenyl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 468.21.73  481

3-Amino-6-(3-sulfamoyl- phenyl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 468.21.74  482

5,6′-Diamino-5′-trifluoromethyl- [2,3′]bipyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 473.11.688 (7.838) 483

5,6′-Diamino-4′-trifluoromethyl- [2,3′]bipyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 473.21.56  484

5,6′-Diamino-2′-fluoro-[2,3′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 423.2 1.69 485

3-Amino-6-(2-amino-4-methoxy- pyrimidin-5-yl)-pyridine-2- carboxylicacid (3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide436.2 1.26  486

3-Amino-6-(2-amino-4- trifluoromethyl-pyrimidin-5-yl)-pyridine-2-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 474.2 1.87  487

6-Phenyl-pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 374.2 2.01  488

3-Amino-6-(2,6-difIuoro-3- isopropoxy-phenyl)-pyridine-2- carboxylicacid (3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide483.2 2.51  489

3-Amino-6-(4-methyl-thiophen- 3-yl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 409.22.11  490

3-Amino-6-(3-cyano-thiophen-2- yl)-pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 420.2 1.96 491

3-Amino-6-(2,6-difluoro-3- propoxy-phenyl)-pyridine-2- carboxylic acid(3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 483.22.70  492

3-Amino-6-(3-ethoxy-2,6- difluoro-phenyl)-pyridine-2- carboxylic acid(3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 469.22.29  493

3-Amino-6-(2,6-difluoro-3- methoxy-phenyl)-pyridine-2- carboxylic acid(3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 455.22.17  494

3-Amino-6-(2,6-difluoro-3- propoxy-phenyl)-5-fluoro-pyridine-2-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 501.2 2.63  495

3-Amino-6-(3-ethoxy-2,6- difluoro-phenyl)-5-fluoro-pyridine-2-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 487.2 2.38  496

3-Amino-6-(2,6-difluoro-3- isopropoxy-phenyl)-5-fluoro-pyridine-2-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 501.2 2.48  497

3-Amino-5-fluoro-6-(3- sulfamoyl-phenyl)-pyridine-2- carboxylic acid(3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 486.11.77  498

3-Amino-5-fluoro-6-(3- methanesulfonylamino-phenyl)-pyridine-2-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 500.2 1.96  499

3-Amino-5-fluoro-6-phenyl- pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 407.1 2.18 500

6-(3-Acetylamino-phenyl)-3- amino-5-fluoro-pyridine-2- carboxylic acid(3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 464.21.90  501

3-Amino-5-fluoro-6-(5-fluoro-2- methoxy-phenyl)-pyridine-2- carboxylicacid (3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide455.2 2.24  502

3-Amino-5-fluoro-6-(2-fluoro-5- methoxy-phenyl)-pyridine-2- carboxylicacid (3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide455.2 2.26  503

3-Amino-6-(2,6-difluoro-3- isopropylcarbamoyl-phenyl)-pyridine-2-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 510.3 2.10  504

5,5′-Diamino-6′-(3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi-pyridinyl-3′-ylcarbamoyl)-3- fluoro-[2,2′]bipyridinyl-6- carboxylic acidmethyl ester 481.2 1.90  505

3-Amino-6-(3-benzyloxy-2,6- difluoro-phenyl)-5-fluoro-pyridine-2-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 549.2 2.80  506

3-Amino-6-(3-butoxy-2,6- difluoro-phenyl)-5-fluoro-pyridine-2-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 515.3 2.83  507

3-Amino-5-fluoro-6-(6-methoxy- pyrazin-2-yl)-pyridine-2- carboxylic acid(3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 439.21.95  508

3-Amino-5-fluoro-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)-pyridine-2-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 510.3 2.14  509

3-Amino-6-(2,6-dimethyl- phenyl)-5-fluoro-pyridine-2- carboxylic acid(3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 435.22.41  510

3-Amino-6-(2,6-difluoro-3- methoxy-phenyl)-5-fluoro-pyridine-2-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 473.2 2.32  511

5-Amino-3-fluoro-2′-methoxy- [2,3′]bipyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 438.21.93  512

5-Amino-3-fluoro-6′-methoxy- [2,2′]bipyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 438.21.89  513

5-Amino-3-fluoro-[2,4′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 408.2 1.24 514

5,6′-Diamino-3-fluoro-[2,3′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 423.2 1.34 515

3-Amino-6-(5- dimethylcarbamoyl-2-fluoro- phenyl)-5-fluoro-pyridine-2-carboxylic acid (3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 496.2 2.14  516

3-Amino-6-(2,6-difluoro-phenyl)- 5-fluoro-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 443.22.25  517

3-Amino-5-fluoro-6-(2-fluoro- phenyl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 425.22.25  518

5-Amino-2-(2,4-difluoro-phenyl)- pyrimidine-4-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 426.2 1.89 519

3-Amino-6-[5-amino-6-(2,2,2- trifluoro-ethoxy)-pyrazin-2-yl]-pyridine-2-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 504.2 2.10  520

3-Amino-6-[5-amino-6-(2- methoxy-ethoxy)-pyrazin-2-yl]-pyridine-2-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 480.2 1.64  521

3-Amino-6-(6- cyclopropylmethoxy-pyrazin-2- yl)-pyridine-2-carboxylicacid (3- amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide461.1 2.38  522

3-Amino-6-(6-hydroxy-pyrazin- 2-yl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 407.11.35  523

3-Amino-6-(6-isobutoxy-pyrazin- 2-yl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 463.22.59  524

3-Amino-6-phenyl-pyridine-2- carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 389.1 2.07  525

6-(4-Piperazin-1-yl-phenyl)- pyridine-2-carboxylic acid [4-(6-amino-2-trifluoromethyl- pyrimidin-4-yl)-pyridin-3-yl]- amide 521.12.20  526

5-Amino-1′,2′,3′,6′-tetrahydro- [2,4′]bipyridinyl-6-carboxylic acid[4-(6-amino-2- trifluoromethyl-pyrimidin-4-yl)- byridin-3-yl]-amide457.2 1.83  527

6-(2,4-Difluoro-phenyl)-pyridine- 2-carboxylic acid [4-(6-amino-2-trifluoromethyl-pyrimidin-4-yl)- pyridin-3-yl]-amide 473.0 3.33  528

6-(2,6-Difluoro-phenyl)-pyridine- 2-carboxylic acid [4-(6-amino-2-trifluoromethyl-pyrimidin-4-yl)- pyridin-3-yl]-amide 473.0 3.16  529

6-(2-Fluoro-phenyl)-pyridine-2- carboxylic acid [4-(6-amino-2-trifluoromethyl-pyrimidin-4-yl)- pyridin-3-yl]-amide 455.1 3.21  530

6-Phenyl-pyridine-2-carboxylic acid [4-(6-amino-2-trifluoromethyl-pyrimidin-4-yl)- pyridin-3-yl]-amide 437.1 3.15  531

3-Amino-6-phenyl-pyridine-2- carboxylic acid [4-(6-amino-2-trifluoromethyl-pyrimidin-4-yl)- pyridin-3-yl]-amide 452.1 3.03  532

3-Amino-6-(2,6-difluoro-3- propoxy-phenyl)-pyridine-2- carboxylic acid[4-(6-amino-2- methylsulfanyl-pyrimidin-4-yl)- pyridin-3-yl]-amide 524.13.33  533

3-Amino-6-(2,6-difluoro-3- propoxy-phenyl)-pyridine-2- carboxylic acid[4-(6-amino-2- methyl-pyrimidin-4-yl)-pyridin- 3-yl]-amide 492.1 2.85 534

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid [4-(6- amino-2-methylsulfanyl- pyrimidin-4-yl)-pyridin-3-yl]- amide533.2  2.70)  535

3-Amino-6-(5- dimethylcarbamoyl-2-fluoro- phenyl)-pyridine-2-carboxylicacid [4-(6-amino-2- methylsulfanyl-pyrimidin-4-yl)- pyridin-3-yl]-amide519.2 2.51  536

3-Amino-6-(2,6-difluoro-3- isopropoxy-phenyl)-pyridine-2- carboxylicacid [4-(6-amino-2- methylsulfanyl-pyrimidin-4-yl)- pyridin-3-yl]-amide524.0 3.26  537

3-Amino-6-(2,6-difluoro-3- isopropoxy-phenyl)-pyridine-2- carboxylicacid [4-(6-amino-2- methyl-pyrimidin-4-yl)-pyridin- 3-yl]-amide 492.22.72  538

3-Amino-6-(2,6-difluoro-3- isopropoxy-phenyl)-pyridine-2- carboxylicacid (2′-amino-6′- methyl-[4,4′]bipyridinyl-3-yl)- amide 491.2 2.57  539

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid (2′- amino-6′-methyl-[4,4′]bi- pyridinyl-3-yl)-amide 500.2 2.15 540

5-Amino-2-(5-ethylcarbamoyl-2- fluoro-phenyl)-pyrimidine-4- carboxylicacid (3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide479.2 1.64  541

5-Amino-2-(2-fluoro-5- isopropoxy-phenyl)-pyrimidine- 4-carboxylic acid(3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 466.32.15  542

5-Amino-2-(2-fluoro-5- isopropylcarbamoyl-phenyl)-pyrimidine-4-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 493.2 1.84  543

5-Amino-2-(2-fluoro-5-methyl- phenyl)-pyrimidine-4-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 422.11.97  544

2-(3-Acetylamino-phenyl)-5- amino-pyrimidine-4-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 447.21.63  545

5-Amino-2-(2,6-difluoro-3- propoxy-phenyl)-pyrimidine-4- carboxylic acid(3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 484.22.19  546

5-Amino-2-(2,3-difluoro-phenyl)- pyrimidine-4-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 426.2 1.85 547

5-Amino-2-(4-chloro-2-fluoro- phenyl)-pyrimidine-4-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 442.12.05  548

5-Amino-2-(5-ethoxy-2-fluoro- phenyl)-pyrimidine-4-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 452.22.02  549

5-Amino-2-(2-fluoro-5-methoxy- phenyl)-pyrimidine-4-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 438.11.84  550

5-Amino-2-(2-fluoro-5-propoxy- phenyl)-pyrimidine-4-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 466.22.25  551

5-Amino-2-(2-methoxy-phenyl)- pyrimidine-4-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 420.1 1.64 552

5-Amino-2′-fluoro-[2,3′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,3′]bipyridinyl-4′-yl)-amide 408.1 1.88 553

6-(2-Fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylic acid[4-(6 amino-2-trifluoromethyl- pyrimidin-4-yl)-pyridin-3-yl]- amide540.3 2.91  554

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid [4-(6- amino-2-trifluoromethyl- pyrimidin-4-yl)-pyridin-3-yl]-amide 555.2 2.83  555

3-Amino-6-(3-ethyl-phenyl)- pyridine-2-carboxylic acid [4-(6-amino-2-methyl-pyrimidin-4-yl)- pyridin-3-yl]-amide 426.1 2.72  556

3-Amino-6-(4-methyl-thiophen- 2-yl)-pyridine-2-carboxylic acid[4-(6-amino-2-methyl-pyrimidin- 4-yl)-pyridin-3-yl]-amide 418.1 2.33 557

3-Amino-6-(2-fluoro-4- trifluoromethyl-phenyl)-pyridine- 2-carboxylicacid [4-(6-amino-2- methyl-pyrimidin-4-yl)-pyridin- 3-yl]-amide 484.22.86  558

3-Amino-6-(2-fluoro-4-methyl- phenyl)-pyridine-2-carboxylic acid[4-(6-amino-2-methyl- pyrimidin-4-yl)-pyridin-3-yl]- amide 430.2 2.57 559

3-Amino-6-phenyl-pyridine-2- carboxylic acid [4-(6-amino-2-methyl-pyrimidin-4-yl)-pyridin- 3-yl]-amide 398.2 2.26  560

3-Amino-5-fluoro-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)-pyridine-2-carboxylic acid [4-(2- amino-6-methyl-pyrimidin-4-yl)-pyridin-3-yl]-amide 519.2 2.70  561

3-Amino-6-(5- dimethylcarbamoyl-2-fluoro- phenyl)-5-fluoro-pyridine-2-carboxylic acid [4-(6-amino-2- methyl-pyrimidin-4-yl)-pyridin-3-yl]-amide 505.1 2.22  562

3-Amino-6-(2,6-difluoro-3- propoxy-phenyl)-pyridine-2- carboxylic acid[4-(2-amino-6- methyl-pyrimidin-4-yl)-pyridin- 3-yl]-amide 492.1 3.11 563

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid [4-(2- amino-6-methyl-pyrimidin-4-yl)- pyridin-3-yl]-amide 501.22.38  564

3-Amino-6-(5- dimethylcarbamoyl-2-fluoro- phenyl)-pyridine-2-carboxylicacid [4-(2-amino-6-methyl- pyrimidin-4-yl)-pyridin-3-yl]- amide 487.22.24  565

3-Amino-6-(2-fluoro-5- propylcarbamoyl-phenyl)- pyridine-2-carboxylicacid [4-(6- amino-2-methyl-pyrimidin-4-yl)- pyridin-3-yl]-amide 501.22.29  566

3-Amino-6-(3-morpholin-4- ylmethyl-phenyl)-pyridine-2- carboxylic acid(3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 488.01.43  567

3-Amino-6-(4-morpholin-4- ylmethyl-phenyl)-pyridine-2- carboxylic acid(3-amino-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 488.01.42  568

3-Fluoro-6-phenyl-pyridine-2- carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide 392.2 1.96  569

3-Fluoro-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)-pyridine-2-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 495.2 2.02  570

6-(2,6-Difluoro-phenyl)-3-fluoro- pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 428.2 2.03 571

3-Fluoro-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 410.2 2.08 572

3-Amino-6-(4-morpholin-4- ylmethyl-phenyl)-pyridine-2- carboxylic acid[4-(6-amino-2- trifluoromethyl-pyrimidin-4-yl)- pyridin-3-yl]-amide551.1 2.10  573

5-Amino-2′-(4-methyl-piperazin- 1-yl)-[2,4′]bipyridinyl-6- carboxylicacid [4-(6-amino-2- trifluoromethyl-pyrimidin-4-yl)- pyridin-3-yl]-amide551.1 1.73  574

3-Amino-6-[4-(4-methyl- piperazin-1-yl)-phenyl]-pyridine- 2-carboxylicacid [4-(6-amino-2- trifluoromethyl-pyrimidin-4-yl)- pyridin-3-yl]-amide550.1 2.14  575

6-(2,6-Difluoro-phenyl)-3-fluoro- pyridine-2-carboxylic acid (5-amino-3-hydroxymethyl-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 458.1 2.10  576

3-Amino-6-(2,6-difluoro-phenyl)- pyridine-2-carboxylic acid (5-amino-3-hydroxymethyl-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 455.1 1.89  577

3-Amino-6-(2-fluoro-5-propoxy- phenyl)-pyridine-2-carboxylic acid(3-amino-5-fluoro-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide483.1 2.53  578

3-Amino-6-phenyl-pyridine-2- carboxylic acid (3-amino-5-fluoro-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 407.1 1.94 579

3-Amino-6-(2-fluoro-5-propoxy- phenyl)-pyridine-2-carboxylic acid(3-amino-5-methoxy- 3,4,5,6-tetrahydro-2H-[1,4′]bi-pyridinyl-3′-yl)-amide 495.2 2.48  580

3-Amino-6-phenyl-pyridine-2- carboxylic acid (3-amino-5-methoxy-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 419.11.97  581

3-Amino-6-(2-fluoro-5-propoxy- phenyl)-pyridine-2-carboxylic acid(3-amino-5-hydroxy-3,4,5,6- tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 481.2 2.45  582

3-Amino-6-phenyl-pyridine-2- carboxylic acid (3-amino-5-hydroxy-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 405.11.86 

Method 14 Example 583 Synthesis of3-amino-6-(2-fluorophenyl)-N-(4-(piperidin-1-yl)pyridin-3-yl)picolinamide

A solution of3-amino-6-bromo-N-(4-(piperidin-1-yl)pyridin-3-yl)picolinamide (1.0 eq),2-fluorophenyl boronic acid (3.0 eq.), Pd(dppf)Cl₂—CH₂Cl₂ (0.15 eq.) in3:1 DME/2M Na₂CO₃ (concentration=0.1 M) was heated at 120° C. withmicrowave irradiation for 1200 seconds. The organic layer was separated,concentrated and directly purified by reverse phase HPLC. Afterlyophilization,(S)-3-amino-N-(4-(3-aminopiperidin-1-yl)pyridin-3-yl)-6-(2-fluoro-4-methylphenyl)picolinamidewas obtained (44%) as the TFA salt. LCMS (m/z): 421.2 (MH⁺); LCR_(t)=2.23 min.

Alternatively, the free base and HCl salt of3-amino-6-(2-fluorophenyl)-N-(4-(piperidin-1-yl)pyridin-3-yl)picolinamidecould be obtained as described in Method 8.

The following compounds were prepared using Method 14. In some cases theanhydrous Suzuki conditions of method 12 (DMF as solvent with 10equivalents of triethylamine) were used.

ID Structure Name MH+ LC 584

5-amino-2′-methoxy-[2,3′]bi- pyridinyl-6-carboxylic acid (4-o-tolyl-pyridin-3-yl)-amide 412.2 3.01 585

3-amino-6-(2,6-difluoro-phenyl)- pyridine-2-carboxylic acid (4-o-tolyl-pyridin-3-yl)-amide 417.1 3.24 586

3-amino-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (4-o-tolyl-pyridin-3-yl)-amide 399.1 3.35 587

5-amino-2′-methoxy-[2,3′]bi- pyridinyl-6-carboxylic acid (3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 405.2 2.83 588

3-amino-6-(2,6-difluoro-phenyl)- pyridine-2-carboxylic acid (3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 410.2 3.14 589

3-amino-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 392.2 3.22 590

3-amino-6-phenyl-pyrazine-2- carboxylic acid (2-oxo-5-piperidin-1-yl-2,3-dihydro-pyrimidin-4-yl)- amide 392.1 2.95 591

3-amino-6-o-tolyl-pyrazine-2- carboxylic acid (2-oxo-5-piperidin-1-yl-2,3-dihydro-pyrimidin-4-yl)- amide 406.2 3.12 592

3-amino-6-(2-fluoro-phenyl)- pyrazine-2-carboxylic acid (2-oxo-5-piperidin-1-yl-2,3-dihydro- pyrimidin-4-yl)-amide 410.1 3.01 593

3-amino-6-(2-trifluoromethyl- phenyl)-pyrazine-2-carboxylic acid(2-oxo-5-piperidin-1-yl-2,3-dihydro- pyrimidin-4-yl)-amide 460.1 3.21594

3-amino-6-(2-methoxy-phenyl)- pyrazine-2-carboxylic acid (2-oxo-5-piperidin-1-yl-2,3-dihydro- pyrimidin-4-yl)-amide 422.1 3 595

3-amino-6-(2-chloro-phenyl)- pyrazine-2-carboxylic acid (2-oxo-5-piperidin-1-yl-2,3-dihydro- pyrimidin-4-yl)-amide 426.1 3.12 596

3-amino-6-(2-trifluoromethoxy- phenyl)-pyrazine-2-carboxylic acid(2-oxo-5-piperidin-1-yl-2,3-dihydro- pyrimidin-4-yl)-amide 476.1 3.34

Method 15 Example 597 Synthesis of3-amino-6-bromo-N-(4-(3-(1-methylpiperidin-4-ylamino)piperidin-1-yl)pyridin-3-yl)pyrazine-2-carboxamide

To a solution of3-amino-N-(4-(3-aminopiperidin-1-yl)pyridin-3-yl)-6-bromopyrazine-2-carboxamidein CH₂Cl₂ at room temperature was added 1-methylpiperidin-4-one (1.5 eq)followed by sodium triacetoxyborohydride (5.0 eq). The reaction wasstirred at room temperature for 12 hours, concentrated, purified byreverse phase HPLC and lyophilized to provide3-amino-6-bromo-N-(4-(3-(1-methylpiperidin-4-ylamino)piperidin-1-yl)pyridin-3-yl)pyrazine-2-carboxamide,(66%). LCMS (m/z): 489.2 (MH⁺).

Following Method 15, the following compounds were prepared.

Example Structure Name MH+ LC 598

3-amino-6-bromo-pyrazine-2- carboxylic acid (3-benzylamino-3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 482.1 2.31 599

3-amino-6-bromo-pyrazine-2- carboxylic acid (3-dibenzylamino-3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 572.2 2.42 600

3-amino-6-bromo-pyrazine-2- carboxylic acid [3-(1-methyl-piperidin-4-ylamino)-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl]-amide 489.2 1.29

Synthesis of trans(+/−)-Benzyl3-(tert-butoxycarbonylamino)-4-hydroxpiperidine-1-carboxylate

Synthesis of trans(+/−)-Benzyl4-(tert-butoxycarbonylamino)-3-hydroxypiperidine-1-carboxylate

A solution of (+/−)benzyl 7-oxa-3-azabicyclo[4.1.0]heptane-3-carboxylate(1.0 equiv.) in saturated ammonium hydroxide aqueous solution andethanol (1:1, 0.05 M solution) in a sealed steel bomb was heated to 70°C. for 5 h. After all volatile materials were removed by N₂ gas stream,ethyl acetate and water were added for work-up. The crude regioisomericmixture, (+/−)benzyl 3-amino-4-hydroxypiperidine-1-carboxylate and(+/−)benzyl 4-amino-3-hydroxypiperidine-1-carboxylate was reacted withBoc₂O (1.0 equiv.) and triethylamine (1.0 equiv.) in dichloromethane(0.1 M solution). After stirred for 2 h at room temperature, thereaction mixture was extracted with dichloromethane. The polar(+/−)-benzyl3-(tert-butoxycarbonylamino)-4-hydroxypiperidine-1-carboxylate andnonpolar (+/−)-benzyl4-(tert-butoxycarbonylamino)-3-hydroxypiperidine-1-carboxylate wereobtained by flash column chromatography (20% to 40% EtOAc in hexanes,28% and 51% each). LCMS (m/z): 351.1 (MH⁺), R_(t)=0.81 min, LCMS (m/z):351.1 (MH⁺), R_(t)=0.83 min.

The enantiomerically pure (3S,4S)-benzyl3-(tert-butoxycarbonylamino)-4-hydroxypiperidine-1-carboxylate and(3R,4R)-benzyl3-(tert-butoxycarbonylamino)-4-hydroxypiperidine-1-carboxylate wereresolved by chiral HPLC (For analysis R_(t)=6.8 min and 9.1 minrespectively; n-heptane:ethanol=70:30 (v:v), Chiralpak AD-H prep 250×4.6mm at 1 mL/min. For preparative separation, n-heptane:ethanol=80:20(v:v), Chiralpak AS 50×500 mm.at 90 mL/min)

Method 16 Synthesis of (+/−)-Benzyl3-(tert-butoxycarbonylamino)-4-(tert-butyldimethylsilyloxy)piperidine-1-carboxylate

To a solution of (+/−)-benzyl3-(tert-butoxycarbonylamino)-4-hydroxypiperidine-1-carboxylate (1.0equiv.) in dichloromethane (0.1 M solution) was added imidazole (1.1equiv.), DMAP (0.1 equiv.), and TBDMSCl (1.1 equiv.) sequentially Thereaction mixture was stirred at room temperature for 20 h. After workedup with dichloromethane, the crude (+/−)-benzyl3-(tert-butoxycarbonylamino)-4-(tert-butyldimethylsilyloxy)piperidine-1-carboxylatewas purified by silica column chromatography (10% to 20% EtOAc inhexanes, 76%). LCMS (m/z): 365.2.

Synthesis of (3R,4R)-benzyl3-(tert-butoxycarbonylamino)-4-(tert-butyldimethylsilyloxy)piperidine-1-carboxylate

Following Method 16, (3R,4R)-benzyl3-(tert-butoxycarbonylamino)-4-hydroxypiperidine-1-carboxylate wasreacted with TBDMSCl, imidazole and DMAP yielding (3R,4R)-benzyl3-(tert-butoxycarbonylamino)-4-(tert-butyldimethylsilyloxy)-piperidine-1-carboxylate.LCMS (m/z): 365.2 (MH⁺); LC R_(t)=6.05 min.

Synthesis of (3S,4S)-benzyl3-(tert-butoxycarbonylamino)-4-(tert-butyldimethylsilyloxy)piperidine-1-carboxylate

Following Method 16, (3S,4S)-benzyl3-(tert-butoxycarbonylamino)-4-hydroxypiperidine-1-carboxylate wasreacted with TBDMSC1, imidazole and DMAP yielding (3S,4S)-benzyl3-(tert-butoxycarbonylamino)-4-(tert-butyldimethylsilyloxy)-piperidine-1-carboxylate.LCMS (m/z): 365.2 (MH⁺); LC R_(t)=6.05 min.

Synthesis of (+/−)-Benzyl4-(tert-butoxycarbonylamino)-3-(tert-butyldimethylsilyloxy)piperidine-1-carboxylate

Following Method 16, (+/−)-benzyl4-(tert-butoxycarbonylamino)-3-hydroxypiperidine-1-carboxylate wasreacted with TBDMSCl, imidazole and DMAP yielding (+/−)-benzyl4-(tert-butoxycarbonylamino)-3-(tert-butyldimethylsilyloxy)-piperidine-1-carboxylate,(81%). LCMS (m/z): 365.2 (MH⁺); LC R_(t)=6.05 min.

Synthesis of (3R,4R)-Benzyl3-(tert-butoxycarbonylamino)-4-fluoropiperidine-1-carboxlate and(3S,4S)-Benzyl-3-(tert-butoxycarbonylamino)-4-fluoropiperidine-1-carboxylate

To a solution of (+/−)-benzyl3-(tert-butoxycarbonylamino)-4-hydroxypiperidine-1-carboxylate (1.0equiv.) in dichloromethane (0.3 M solution) was added DAST at −78° C.The reaction mixture was slowly warmed up to room temperature for 15 h.After quenched with saturated sodium bicarbonate aqueous solution, ethylacetate and water were added for work-up. The (+/−)-benzyl3-(tert-butoxycarbonylamino)-4-fluoropiperidine-1-carboxylate wasobtained by silica column chromatography (30% EtOAc in hexanes, 40%).LCMS (m/z): 253.1; LC R_(t)=4.08 min. The enantiomerically pure(3R,4R)-benzyl3-(tert-butoxycarbonylamino)-4-fluoropiperidine-1-carboxylate and(3S,4S)-benzyl3-(tert-butoxycarbonylamino)-4-fluoropiperidine-1-carboxylate wereresolved by chiral HPLC (for analysis: R_(t)=9.4 min and 12.6 minrespectively; n-heptane:isopropanol=90:10 (v:v), Chiralpak AS 250×4.6 mmat 1 mL/min. For preparative separation, n-heptane:isopropanol=90:10(v:v), Chiralpak AS 50×500 mm.at 90 mL/min)

Synthesis of trans-(+/−)-Benzyl4-fluoro-3-hydroxypiperidine-1-carboxylate

A solution of (+/−)-benzyl7-oxa-3-azabicyclo[4.1.0]heptane-3-carboxylate (1.0 equiv.) and Et₃N.3HF(1 equiv.) in a sealed glass flask were heated to 100° C. for 15 h. Thereaction mixture was extracted with ethyl acetate, which was washed withwater and brine. The organic phase was dried over anhydrous sodiumsulfate and filtered. After volatile materials were removed,(+/−)-benzyl 4-fluoro-3-hydroxypiperidine-1-carboxylate was obtained bysilica column chromatography (20% to 40% EtOAc in hexanes, 53%). LCMS(m/z): 254.1 (MH⁺); LC R_(t)=2.86 min.

Synthesis of trans (+/−)-Benzyl3-(1,3-dioxoisoindolin-2-yl)-4-fluoropiperidine-1-carboxylate

To a solution of triphenylphosphine (3.0 equiv.) in toluene (0.25 Msolution) was added DEAD (3.0 equiv.) at room temperature, which wasstirred for 15 min. Then, (+/−)-benzyl4-fluoro-3-hydroxypiperidine-1-carboxylate (1.0 equiv.) was added to thereaction mixture. After being stirred for 10 min, phthalimide (3.0equiv.) was added and the reaction mixture was stirred for 15 h. Thereaction mixture was extracted with ethyl acetate, which was washed withwater and brine. The organic phase was dried over anhydrous sodiumsulfate and filtered. After volatile materials were removed,(+/−)-benzyl3-(1,3-dioxoisoindolin-2-yl)-4-fluoropiperidine-1-carboxylate wasobtained by silica column chromatography (10% to 20% EtOAc in hexanes,20%). LCMS (m/z): 383.0 (MH⁺), R_(t)=1.0 min.

Synthesis of (+/−)-Benzyl3-(tert-butoxycarbonylamino)-4-oxopiperidine-1-carboxylate

To a solution of (+/−)-benzyl3-(tert-butoxycarbonylamino)-4-hydroxypiperidine-1-carboxylate indichloromethane (0.1 M solution) was added Dess-Martin periodinane (1.5equiv.). The reaction mixture was stirred at room temperature for 16 h.Then, the saturated NaHCO₃ and 0.1 N Na₂S₂O₃ aqueous solutions wereadded to the reaction mixture, which was stirred for 30 min and workedup with ethyl acetate. The crude benzyl3-(tert-butoxycarbonylamino)-4-oxopiperidine-1-carboxylate was purifiedby flash chromatography (30% EtOAc in hexanes, 70%). LCMS (m/z): 249.2(MH⁺); LC R_(t)=3.98 min. ¹H NMR (CDCl₃): 1.41 (1H, s), 2.1-2.59 (1H,m), 2.73 (1H, m), 3.09 (1H, m), 4.30 (1H, m), 4.52 (1H, m), 4.90 (1H,m), 5.19 (2H, m), 5.45 (1H, m), 7.39 (5H, m).

Synthesis of (+/−)-Benzyl3-(tert-butoxycarbonylamino)-4,4-difluoropiperidine-1-carboxylate

To a solution of (+/−)-benzyl3-(tert-butoxycarbonylamino)-4-oxopiperidine-1-carboxylate indichloromethane (0.3 M solution) was added DAST (3.0 equiv.). Thereaction mixture was stirred at room temperature for 20 h. Afterquenched with saturated NaHCO₃ aqueous solution until no bubbling, thereaction mixture was extracted with CH₂Cl₂. The crude (+/−)-benzyl3-(tert-butoxycarbonylamino)-4,4-difluoropiperidine-1-carboxylate waspurified by flash chromatography (10% to 40% EtOAc in hexanes, 35%).LCMS (m/z): 271.0 (MH⁺), LC R_(t)=4.2 min. ¹H NMR (CDCl₃): 1.26 (9H, s),1.90 (1H, m), 2.11 (1H, m), 2.98 (1H, t, J=11.2 Hz), 3.20 (1H, t,J=11.6), 4.00 (1H, m), 4.13 (1H, m), 4.76 (1H, m), 5.11 (1H, m), 7.36(1H, m).

Method 17 Synthesis ofcis-(+/−)-2-(4-fluoropiperidin-3-yl)isoindoline-1,3-dione

To a solution of (+/−)-benzyl3-(1,3-dioxoisoindolin-2-yl)-4-fluoropiperidine-1-carboxylate (1.0equiv.) in ethanol and ethyl acetate (1:1, 0.2 M solution) was addedPd/C (20 wt %) under N₂ atmosphere. The reaction mixture was flushedwith H₂ gas, equipped with H₂ gas balloon, and stirred for 16 h at roomtemperature. The reaction mixture was filtered through Celite® Pad andthe filtrate was dried in vacuo. The crude(+/−)-2-(4-fluoropiperidin-3-yl)isoindoline-1,3-dione was used for thenext step without further purification (>99%). LCMS (m/z): 249.1 (MH⁺),R_(t)=0.49 min.

Synthesis of trans-(+/−)-tert-Butyl 4-fluoropiperidin-3-ylcarbamate

Method 17 was followed using (+/−)trans-benzyl3-(tert-butoxycarbonylamino)-4-fluoropiperidine-1-carboxylate (1.0equiv.) yielding crude (+/−)-trans tert-butyl4-fluoropiperidin-3-ylcarbamate (93%). LCMS (m/z): 219.2 (MH⁺), LCR_(t)=0.45 min.

Synthesis of tert-butyl (3R,4R)-4-fluoropiperidin-3-ylcarbamate

Method 17 was followed using (3R,4R)-benzyl3-(tert-butoxycarbonylamino)-4-fluoropiperidine-1-carboxylate (1.0equiv.) yielding crude (+/−)-tert-butyl4-fluoropiperidin-3-ylcarbamate(93%). LCMS (m/z): 219.2 (MH⁺), LCR_(t)=0.45 min.

Synthesis of tert-butyl (3S,4S)-4-fluoropiperidin-3-ylcarbamate

Method 17 was followed using (3S,4S)-benzyl3-(tert-butoxycarbonylamino)-4-fluoropiperidine-1-carboxylate (1.0equiv.) yielding crude (+/−)-tert-butyl 4-fluoropiperidin-3-ylcarbamate(93%). LCMS (m/z): 219.2 (MH⁺), LC R_(t)=0.45 min.

Synthesis of trans-(+/−)-Butyl4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate

Method 17 was followed using (+/−)-benzyl3-(tert-butoxycarbonylamino)-4-(tert-butyldimethylsilyloxy)piperidine-1-carboxylate(1.0 equiv.) yielding crude (+/−)-butyl4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate (>99%). LCMS(m/z): 331.3 (MH⁺).

Synthesis of tert-butyl(3R,4R)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate

Method 17 was followed using (3R,4R)-benzyl3-(tert-butoxycarbonylamino)-4-(tert-butyldimethylsilyloxy)piperidine-1-carboxylate(1.0 equiv.) yielding crude tert-butyl(3R,4R)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate (>99%).LCMS (m/z): 331.3 (MH⁺).

Synthesis of tert-butyl(3R,4R)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate

Method 17 was followed using (3S,4S)-benzyl3-(tert-butoxycarbonylamino)-4-(tert-butyldimethylsilyloxy)piperidine-1-carboxylate(1.0 equiv.) yielding crude tert-butyl(3S,4S)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate (>99%).LCMS (m/z): 331.3 (MH⁺).

Synthesis of trans-(+/−)-tert-Butyl3-(tert-butyldimethylsilyloxy)piperidin-4-ylcarbamate

Method 17 was followed using (+/−)-benzyl4-(tert-butoxycarbonylamino)-3-hydroxypiperidine-1-carboxylate (1.0equiv.) yielding (+/−)-tert-butyl3-(tert-butyldimethylsilyloxy)piperidin-4-ylcarbamate (>99%). LCMS(m/z): 331.2 (MH⁺).

Synthesis of (+/−)-tert-Butyl 4,4-difluoropiperidin-3-ylcarbamate

Method 17 was followed using (+/−)-benzyl3-(tert-butoxycarbonylamino)-4,4-difluoropiperidine-1-carboxylate (1.0equiv.) yielding crude (+/−)-tert-butyl4,4-difluoropiperidin-3-ylcarbamate, (>99%). LCMS (m/z): 237.0 (MH⁺).

Synthesis of trans-(+/−)-tert-Butyl4-(tert-butyldimethylsilyloxy)-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate

Method 1 of Example 1 was followed using 1 eq each of4-chloro-3-nitropyidine, trans-(+/−)-butyl4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate and triethylaminein DMF yielding (+/−)-tert-butyl4-(tert-butyldimethylsilyloxy)-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate,(98%). LCMS (m/z): 453.3 (MH⁺); LC R_(t)=4.01 min.

Synthesis of tert-butyl(3R,4R)-4-(tert-butyldimethylsilyloxy)-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate

Method 1 of Example 1 was followed using 1 eq each of4-chloro-3-nitropyidine, tert-butyl(3R,4R)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate andtriethylamine in DMF yielding tert-butyl(3R,4R)-4-(tert-butyldimethylsilyloxy)-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate(98%). LCMS (m/z): 453.3 (MH⁺); LC R_(t)=4.01 min.

Synthesis of tert-butyl(3S,4S)-4-(tert-butyldimethylsilyloxy)-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate

Method 1 of Example 1 was followed using 1 eq each of4-chloro-3-nitropyidine, tert-butyl(3S,4S)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate andtriethylamine in DMF yielding tert-butyl(3S,4S)-4-(tert-butyldimethylsilyloxy)-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate(98%). LCMS (m/z): 453.3 (MH⁺); LC R_(t)=4.01 min.

Synthesis of trans-(+/−)-tert-butyl3-(tert-butyldimethylsilyloxy)-1-(3-nitropyridin-4-yl)piperidin-4-ylcarbamate

Method 1 of Example 1 was followed using 1 eq each of4-chloro-3-nitropyidine, (+/−)-tert-butyl3-(tert-butyldimethylsilyloxy)piperidin-4-ylcarbamate and triethylaminein ethanol yielding (+/−)-tert-butyl3-(tert-butyldimethylsilyloxy)-1-(3-nitropyridin-4-yl)piperidin-4-ylcarbamate,(75%). LCMS (m/z): 453.2 (MH⁺); LC R_(t)=3.46 min.

Synthesis of trans-(+/−)-tert-Butyl4-fluoro-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate

Method 1 of Example 1 was followed using 1 eq each of4-chloro-3-nitropyidine, (+/−)-tert-butyl4-fluoropiperidin-3-ylcarbamate and triethylamine in ethanol yielding(+/−)-tert-butyl4-fluoro-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate, (91%). LCMS(m/z): 341.0 (MH⁺); LC R_(t)=2.37 min.

Synthesis of tert-butyl(3R,4R)-4-fluoro-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate

Method 1 of Example 1 was followed using 1 eq each of4-chloro-3-nitropyidine, tert-butyl(3R,4R)-4-fluoropiperidin-3-ylcarbamate and triethylamine in ethanolyielding tert-butyl(3R,4R)-4-fluoro-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate, (91%).LCMS (m/z): 341.0 (MH⁺); LC R_(t)=2.37 min.

Synthesis of tert-butyl(3S,4S)-4-fluoro-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate

Method 1 of Example 1 was followed using 1 eq each of4-chloro-3-nitropyidine, tert-butyl(3S,4S)-4-fluoropiperidin-3-ylcarbamate and triethylamine in ethanolyielding tert-butyl(3S,4S)-4-fluoro-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate, (91%).LCMS (m/z): 341.0 (MH⁺); LC R_(t)=2.37 min.

Synthesis of (+/−)-tert-Butyl4,4-difluoro-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate

Method 1 of Example 1 was followed using 1 eq each of4-chloro-3-nitropyidine, (+/−)-tert-butyl4,4-difluoropiperidin-3-ylcarbamate and triethylamine in ethanolyielding (+/−)-tert-butyl4,4-difluoro-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate, (91%). LCMS(m/z): 359.1 (MH⁺).

Synthesis ofcis-(+/−)-2-(4-fluoro-1-(3-nitropyridin-4-yl)piperidin-3-yl)isoindoline-1,3-dione

Method 1 of Example 1 was followed using 1 eq each of4-chloro-3-nitropyidine,(+/−)-2-(4-fluoropiperidin-3-yl)isoindoline-1,3-dione and triethylaminein DMF yielding(+/−)-2-(4-fluoro-1-(3-nitropyridin-4-yl)piperidin-3-yl)isoindoline-1,3-dione,(45%). LCMS (m/z): 371.1 (MH⁺); LC R_(t)=2.23 min.

Synthesis of trans-(+/−)-tert-Butyl1-(3-aminopyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate

Following Method 2 of Example 49, (+/−)-tert-butyl4-(tert-butyldimethylsilyloxy)-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamatein ethanol and ethyl acetate (1:1, 0.1 M solution) was reduced yielding(+/−)-tert-butyl1-(3-aminopyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate,(>99%). LCMS (m/z): 423.2 (MH⁺); LC R_(t)=3.78 min.

Synthesis of tert-butyl(3R,4R)-1-(3-aminopyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate

Following Method 2 of Example 49, tert-butyl(3R,4R)-4-(tert-butyldimethylsilyloxy)-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamatein ethanol and ethyl acetate (1:1, 0.1 M solution) was reduced yieldingtert-butyl(3R,4R)-1-(3-aminopyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate,(>99%). LCMS (m/z): 423.2 (MH⁺); LC R_(t)=3.78 min.

Synthesis of tert-butyl(3S,4S)-1-(3-aminopyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate

Following Method 2 of Example 49, tert-butyl(3R,4R)-4-(tert-butyldimethylsilyloxy)-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamatein ethanol and ethyl acetate (1:1, 0.1 M solution) was reduced yieldingtert-butyl(3R,4R)-1-(3-aminopyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate,(>99%). LCMS (m/z): 423.2 (MH⁺); LC R_(t)=3.78 min.

Synthesis of trans-(+/−)-tert-Butyl1-(3-aminopyridin-4-yl)-3-(tert-butyldimethylsilyloxy)piperidin-4-ylcarbamate

Following Method 2 of Example 49, (+/−)-tert-butyl3-(tert-butyldimethylsilyloxy)-1-(3-nitropyridin-4-yl)piperidin-4-ylcarbamatein ethanol and ethyl acetate (1:1, 0.1 M solution) was reduced yielding(+/−)-tert-butyl1-(3-aminopyridin-4-yl)-3-(tert-butyldimethylsilyloxy)piperidin-4-ylcarbamate,(>99%). LCMS (m/z): 423.3 (MH⁺); LC R_(t)=3.62 min.

Synthesis of trans-(+/−)-tert-butyl1-(3-aminopyridin-4-yl)-4-fluoropiperidin-3-ylcarbamate

Following Method 2 of Example 49, (+/−)-tert-butyl4-fluoro-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate in ethanol andethyl acetate (1:1, 0.1 M solution) was reduced yielding(+/−)-tert-butyl1-(3-aminopyridin-4-yl)-4-fluoropiperidin-3-ylcarbamate, (>99%). LCMS(m/z): 311.2 (MH⁺); LC R_(t)=2.14 min.

Synthesis of tert-butyl(3R,4R)-1-(3-aminopyridin-4-yl)-4-fluoropiperidin-3-ylcarbamate

Following Method 2 of Example 49, tert-butyl(3R,4R)-4-fluoro-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate inethanol and ethyl acetate (1:1, 0.1 M solution) was reduced yieldingtert-butyl(3R,4R)-1-(3-aminopyridin-4-yl)-4-fluoropiperidin-3-ylcarbamate, (>99%).LCMS (m/z): 311.2 (MH⁺); LC R_(t)=2.14 min.

Synthesis of tert-butyl(3S,4S)-1-(3-aminopyridin-4-yl)-4-fluoropiperidin-3-ylcarbamate

Following Method 2 of Example 49, tert-butyl(3S,4S)-4-fluoro-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate inethanol and ethyl acetate (1:1, 0.1 M solution) was reduced yieldingtert-butyl(3R,4R)-1-(3-aminopyridin-4-yl)-4-fluoropiperidin-3-ylcarbamate, (>99%).LCMS (m/z): 311.2 (MH⁺); LC R_(t)=2.14 min.

Synthesis of (+/−)-tert-Butyl1-(3-aminopyridin-4-yl)-4,4-difluoropiperidin-3-ylcarbamate

Following Method 2 of Example 49, (+/−)-tert-butyl4,4-difluoro-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate andtriethylamine in ethanol and ethyl acetate (1:1, 0.1 M solution) wasreduced yielding (+/−)-tert-butyl1-(3-aminopyridin-4-yl)-4,4-difluoropiperidin-3-ylcarbamate, (>99%).LCMS (m/z): 329.1 (MH⁺).

Synthesis ofcis-(+/−)-2-(1-(3-aminopyridin-4-yl)-4-fluoropiperidin-3-yl)isoindoline-1,3-dione

Following Method 2 of Example 49,(+/−)-2-(4-fluoro-1-(3-nitropyridin-4-yl)piperidin-3-yl)isoindoline-1,3-dionein ethanol and ethyl acetate (1:1, 0.1 M solution) was reduced yielding(+/+2-(1-(3-aminopyridin-4-yl)-4-fluoropiperidin-3-yl)isoindoline-1,3-dione,(87%). LCMS (m/z): 341.1 (MH⁺); LC R_(t)=2.23 min.

Method 18 Synthesis of 3-methylpiperidine-3-carboxylic acid

TFA was added to a solution of1-(tert-butoxycarbonyl)-3-methylpiperidine-3-carboxylic acid (1 eq) inCH₂Cl₂ (0.5 M). After stirring for 3 h at rt the reaction mixture wasconcentrated in vacuo and azeotroped once with toluene to give3-methylpiperidine-3-carboxylic acid (TFA salt). The crude product wasused for the next step without further purification.

Method 19 Synthesis of3-methyl-1-(3-nitropyridin-4-yl)piperidine-3-carboxylic acid

4-chloro-3-nitropyridine (1.1 eq) was added to a solution of3-methylpiperidine-3-carboxylic acid (1 eq) and DIEA (3 eq) in iPrOH(0.1 M). The reaction mixture was heated in a 60° C. oil bath for 3 hthen concentrated in vacuo. The crude residue was diluted with EtOAc andwashed with 1.0 N NaOH. The combined aqueous washes were acidified topH=4 with 1.0 N HCl and extracted with CH₂Cl₂. The combined organicphases were dried over anhydrous MgSO₄, filtered, and concentrated invacuo to give 3-methyl-(3-nitropyridin-4-yl)piperidine-3-carboxylicacid. LC/MS (m/z): 266.2 (MH⁺).

Synthesis of tert-butyl3-methyl-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate

Diphenylphosphoryl azide (1.2 eq) was added to a mixture of3-methyl-(3-nitropyridin-4-yl)piperidine-3-carboxylic acid (1 eq) andanhydrous tBuOH (0.3 M), followed shortly by Et₃N (2 eq). The reactionflask was fitted with an air-cooled reflux condenser and bubble vent,then heated in an 85° C. oil bath for 3 days. The crude mixture wasdiluted with EtOAc, washed with brine, then dried over anhydrous MgSO₄,filtered, and concentrated in vacuo. The crude residue was dissolved inCH₂Cl₂, loaded onto a SiO₂ column, and purified by flash chromatography(10-20-40% EtOAc in hexanes) to give tert-butyl3-methyl-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate. LC/MS (m/z):337.2 (MH⁺).

Synthesis of tert-butyl1-(3-aminopyridin-4-yl)-3-methylpiperidin-3-ylcarbamate

10% Palladium on carbon (0.1 eq) was added to a N₂-flushed solution oftert-butyl 3-methyl-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate (1eq) in MeOH (0.2 M). The reaction was purged with H₂ under atmosphericpressure for 16 h at rt. The crude solids were filtered through a pad ofCelite on a paper lined Buchner funnel, washed with EtOAc and MeOH, thenconcentrated in vacuo. The crude residue was dissolved in CH₂Cl₂, loadedonto a SiO₂ column, and purified by flash chromatography (1:2 EtOAc inhexanes+5% MeOH) to give tert-butyl1-(3-aminopyridin-4-yl)-3-methylpiperidin-3-ylcarbamate. LC/MS (m/z):307.2 (MH⁺).

Synthesis of tert-butyl3-(trifluoromethyl)-3-(trimethylsilyloxy)piperidine-1-carboxylate

To an ice-bath cooled solution of tert-butyl3-oxopiperidine-1-carboxylate (1 eq) in THF (0.4 M) was addedtrimethyl(trifluoromethyl)silane (1.2 eq) followed shortly by a 1.0Msolution of TBAF in THF (0.05 eq). The reaction was warmed to rt andstirred for 4 h, then concentrated in vacuo. The resulting residue wasdissolved in EtOAc, washed with brine, then dried over anhydrous MgSO₄,filtered, and concentrated in vacuo to give tert-butyl3-(trifluoromethyl)-3-(trimethylsilyloxy)piperidine-1-carboxylate, whichwas carried on crude and used without further purification.

Synthesis of tert-butyl3-hydroxy-3-(trifluoromethyl)piperidine-1-carboxylate

A 1.0 M solution of TBAF in THF (1 eq) was added to a solution oftert-butyl3-(trifluoromethyl)-3-(trimethylsilyloxy)piperidine-1-carboxylate (1 eq)in THF (0.2 M). After stirring for 16 h at rt the reaction mixture wasconcentrated in vacuo. The resulting residue was dissolved in EtOAc,washed with brine, then dried over anhydrous MgSO₄, filtered, andconcentrated in vacuo. The crude residue was dissolved in CH₂Cl₂, loadedonto a SiO₂ column, and purified by flash chromatography to givetert-butyl 3-hydroxy-3-(trifluoromethyl)piperidine-1-carboxylate. ¹⁹FNMR (282 MHz, CDCl₃): 6-83.74 ppm; ¹H NMR (300 MHz, CDCl₃): δ 4.04-4.16(m, 2H), 2.72-3.01 (m, 2H), 1.50-2.04 (m, 4H), 1.47 (s, 9H).

Synthesis of 3-(trifluoromethyl)piperidin-3-ol

Method 18 was followed using tert-butyl3-hydroxy-3-(trifluoromethyl)piperidine-1-carboxylate, yielding3-(trifluoromethyl)piperidin-3-ol (TFA salt). The crude product was usedfor the next step without further purification.

Synthesis of 1-(3-nitropyridin-4-yl)-3-(trifluoromethyl)piperidin-3-ol

Method 19 was followed using 3-(trifluoromethyl)piperidin-3-ol, yielding1-(3-nitropyridin-4-yl)-3-(trifluoromethyl)piperidin-3-ol. LC/MS (m/z):292.0 (MH⁺).

Synthesis of 1-(3-aminopyridin-4-yl)-3-(trifluoromethyl)piperidin-3-ol

Method 2 of example 49 was followed using1-(3-nitropyridin-4-yl)-3-(trifluoromethyl)piperidin-3-ol, yielding1-(3-aminopyridin-4-yl)-3-(trifluoromethyl)piperidin-3-ol. LC/MS (m/z):262.0 (MH⁺).

Synthesis of tert-butyl(3S,5R)-5-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate

tert-Butyl (3S,5R)-5-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamatewas prepared according to the patent procedure as described by Y, Zhou;WO2005028467.

Synthesis of tert-butyl(3S,5R)-5-(tert-butyldimethylsilyloxy)-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate

Method 19 was followed was followed using tert-Butyl(3S,5R)-5-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate, yieldingtert-butyl(3S,5R)-5-(tert-butyldimethylsilyloxy)-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate.LC/MS (m/z): 453.2 (MH⁺).

Synthesis of tert-butyl(3S,5R)-1-(3-aminopyridin-4-yl)-5-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate

Method 2 was followed using tert-butyl(3S,5R)-5-(tert-butyldimethylsilyloxy)-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate,yielding tert-butyl(3S,5R)-1-(3-aminopyridin-4-yl)-5-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate.LC/MS (m/z): 423.2 (MH⁺).

Synthesiscis-(+/−)-1-(benzyloxycarbonyl)-5-(tert-butoxycarbonylamino)piperidine-3-carboxylicacid

To a solution ofcis-(+/−)-5-(tert-butoxycarbonylamino)piperidine-3-carboxylic acid (1.0eq.) in dichloromethane (0.2 M) was added DIEA(1.1 eq.), followed byN-(benzyloxycarbonyloxy)succinimide (1.0 eq.); the reaction was stirredat r.t. overnight. The solvent was removed under reduced pressure. Tothe crude was added EtOAc and 1N HCl. After extraction, the organiclayer was washed with brine, dried and filtered, and concentrated toyieldcis-(+/−)-1-(benzyloxycarbonyl)-5-(tert-butoxycarbonylamino)piperidine-3-carboxylicacid (99% yield) LCMS (m/z): 379.2 (MH⁺); LC R_(t)=3.55 min.

Synthesis of cis-(+/−)-benzyl3,5-bis(tert-butoxycarbonylamino)piperidine-1-carboxylate

To a solution ofcis-(+/−)-1-(benzyloxycarbonyl)-5-(tert-butoxycarbonylamino)piperidine-3-carboxylic acid (1.2 g, 3.17 mmol), DPPA(Diphenylphosphorylazide, 1.04 g, 3.81 mmol) and DIEA(1.1 mL, 6.35 mmol) in t-BuOH(10 mL)was heated to 90° C. over night. The solvent was removed under reducedpressure. To the crude was added EtOAc (300 mL), the organic layer waswashed with saturated NaHCO₃(150 mL) and brine, dried and filtered, andconcentrated to give the crude. The crude material was further purifiedby silica gel chromatography to yielding cis-(+/−)-benzyl3,5-bis(tert-butoxycarbonylamino)piperidine-1-carboxylate, (23%). LCMS(m/z): 350(minus one Boc(MH⁺); LC R_(t)=4.40 min.

Synthesis of tert-butyl cis-(+/−)-piperidine-3,5-diyldicarbamate

Method 17 was followed using cis-(+/−)-benzyl3,5-bis(tert-butoxycarbonylamino)piperidine-1-carboxylate yieldingtert-butyl cis-(+/−)-piperidine-3,5-diyldicarbamate, (% yield 99%). LCMS(m/z): 316.2 (MH⁺).

Synthesis of tert-butylcis-(+/−)-1-(3-nitropyridin-4-yl)piperidine-3,5-diyldicarbamate

Method 1 of Example 1 was followed using 1 equivalent each of4-chloro-3-nitropyridine, tert-butylcis-(+/−)-piperidine-3,5-diyldicarbamate and triethylamine in DMFyielding tert-butylcis-(+/−)-1-(3-nitropyridin-4-yl)piperidine-3,5-diyldicarbamate, LCMS(m/z): 438.2 (MH⁺); LC R_(t)=2.95 min.

Synthesis ofcis-tert-butyl(+/−)-1-(3-aminopyridin-4-yl)piperidine-3,5-diyldicarbamate

Following Method 2 of Example 49,cis-(+/−)1-(3-nitropyridin-4-yl)piperidine-3,5-diyldicarbamate inethanol was reduced yieldingcis-tert-butyl(+/−)-1-(3-aminopyridin-4-yl)piperidine-3,5-diyldicarbamate,(78%). LCMS (m/z): 408.2 (MH⁺); LC R_(t)=2.63 min.

Synthesis of (S)-1-(3-nitropyridin-4-yl)piperidin-3-ol

Method 1 of Example 1 was followed using 1 equivalent each of4-chloro-3-nitropyridine, (S)-3-hydroxypiperidine and triethylamine inDMF yielding (S)-1-(3-nitropyridin-4-yl)piperidin-3-ol, LCMS (m/z):224.1 (MH⁺); LC R_(t)=1.06 min.

Synthesis of (R)-1-(3-nitropyridin-4-yl)piperidin-3-ol

Method 1 of Example 1 was followed using 1 equivalent each of4-chloro-3-nitropyridine, (R)-3-hydroxypiperidine and triethylamine inDMF yielding (R)-1-(3-nitropyridin-4-yl)piperidin-3-ol, LCMS (m/z):224.1 (MH⁺); LC R_(t)=1.06 min.

Synthesis of (+/−)-1-(3-nitropyridin-4-yl)piperidin-3-ol

Method 1 of Example 1 was followed using 1 equivalent each of4-chloro-3-nitropyridine, (+/−)-3-hydroxypiperidine and triethylamine inDMF yielding (+/−)-1-(3-nitropyridin-4-yl)piperidin-3-ol, LCMS (m/z):224.1 (MH⁺); LC R_(t)=1.06 min.

Synthesis of(S)-4-(3-(tert-butyldimethylsilyloxy)piperidin-1-yl)-3-nitropyridine

To a solution of (S)-1-(3-nitropyridin-4-yl)piperidin-3-ol and TBDMSCl(2.1 equiv.) in DMF was added imidazole (4 equiv.). The reaction washeated to 50° C. overnight. The reaction was dissolved in EtOAC andwashed with water followed by saturate brine, dried and filtered, andconcentrated to give the crude. The crude material was further purifiedby silica gel chromatography to yield desired product

-   (S)-4-(3-(tert-butyl    dimethylsilyloxy)piperidin-1-yl)-3-nitropyridine. LCMS (m/z): 338.2    (MH⁺); LC R_(t)=3.43 min.

Synthesis of(R)-4-(3-(tert-butyldimethylsilyloxy)piperidin-1-yl)-3-nitopyridine

To a solution of (R)-1-(3-nitropyridin-4-yl)piperidin-3-ol and TBDMSCl(2.1 equiv.) in DMF was added imidazole (4 equiv.). The reaction washeated to 50° C. over night. The reaction was dissolved in EtOAc andwashed with water followed by saturate brine, dried and filtered, andconcentrated to give the crude. The crude material was further purifiedby silica gel chromatography to yield desired product(R)-4-(3-(tert-butyl dimethylsilyloxy)piperidin-1-yl)-3-nitropyridine.LCMS (m/z): 338.2 (MH⁺); LC R_(t)=3.43 min.

Synthesis of(+/−)-4-(3-(tert-butyldimethylsilyoxy)piperidin-1-yl)-3-nitropyridine

To a solution of (+/−)-1-(3-nitropyridin-4-yl)piperidin-3-ol and TBDMSCl(2.1 equiv.) in DMF was added imidazole (4 equiv.). The reaction washeated to 50° C. over night. The reaction was dissolved in EtOAc andwashed with water followed by brine, dried and filtered, andconcentrated to give the crude. The crude material was further purifiedby silica gel chromatography to yield desired product(+/−)-4-(3-(tert-butyl dimethylsilyloxy)piperidin-1-yl)-3-nitropyridine.LCMS (m/z): 338.2 (MH⁺); LC R_(t)=3.43 min.

Synthesis of(+/−)-4-(3-(tert-butyldimethylsilyloxy)piperidin-1-yl)pyridin-3-amine

Following Method 2 of Example 49, (+/−)-4-(3-(tert-butyldimethylsilyloxy)piperidin-1-yl)-3-nitropyridine in ethanol was reducedyieldingtert-butyl(+/−)-4-(3-(tert-butyldimethylsilyloxy)piperidin-1-yl)pyridin-3-amine.LCMS (m/z): 308.2 (MH⁺); LC R_(t)=3.47 min.

Synthesis of(S)-4-(3-(tert-butyldimethylsilyloxy)piperidin-1-yl)pyridin-3-amine

Following Method 2 of Example 49, (S)-4-(3-(tert-butyldimethylsilyloxy)piperidin-1-yl)-3-nitropyridine in ethanol was reducedyieldingtert-butyl(S)-4-(3-(tert-butyldimethylsilyloxy)piperidin-1-yl)pyridin-3-amine(67% yield 3 steps). LCMS (m/z): 308.2 (MH⁺); LC R_(t)=3.47 min.

Synthesis of(R)-4-(3-(tert-butyldimethylsilyloxy)piperidin-1-yl)pyridin-3-amine

Following Method 2 of Example 49, (R)-4-(3-(tert-butyldimethylsilyloxy)piperidin-1-yl)-3-nitropyridine in ethanol was reducedyielding tert-butyl

-   (S)-4-(3-(tert-butyldimethylsilyloxy)piperidin-1-yl)pyridin-3-amine    LCMS (m/z): 308.2 (MH⁺); LC R_(t)=3.47 min.

Synthesis of3-amino-6-bromo-N(4-(3-(tert-butyldimethylsilyoxy)piperidin-1-yl)pyridin-3-yl)picolinamide

Following Method 11 of Example 305,tert-butyl(+/−)-4-(3-(tert-butyldimethylsilyloxy)piperidin-1-yl)pyridin-3-aminewas coupled to 3-amino-6-bromopicolinic acid yielding3-amino-6-bromo-N-(4-(3-(tert-butyldimethylsilyloxy)piperidin-1-yl)pyridin-3-yl)picolinamide.LCMS (m/z): 506.2 (MH⁺); LC R_(t)=4.03 min.

Synthesis of3-amino-6-bromo-N-(4-(3-(tert-butyldimethylsilyoxy)piperidin-1-yl)pyridin-3-yl)picolinamide

Following Method 11 of Example 305,tert-butyl(+/−)-4-(3-(tert-butyldimethylsilyloxy)piperidin-1-yl)pyridin-3-aminewas coupled to 3-amino-6-bromopicolinic acid yielding3-amino-6-bromo-N-(4-(3-(tert-butyldimethylsilyloxy)-piperidin-1-yl)pyridin-3-yl)picolinamide.LCMS (m/z): 506.2 (MH⁺); LC R_(t)=4.03 min.

Synthesis(S)-3-amino-6-bromo-N-(4-(3-(tert-butyldimethylsilyoxy)piperidin-1-yl)pyridin-3-yl)picolinamide

Following Method 11 of Example 305,tert-butyl(S)-4-(3-(tert-butyldimethylsilyloxy)piperidin-1-yl)pyridin-3-aminewas coupled to 3-amino-6-bromopicolinic acid yielding(S)-3-amino-6-bromo-N-(4-(3-(tert-butyldimethyl-silyloxy)piperidin-1-yl)pyridin-3-yl)picolinamide.LCMS (m/z): 506.2 (MH⁺); LC R_(t)=4.03 min.

Synthesis of(R)-3-amino-6-bromo-N-(4-(3-(tert-butyldimethylsilyloxy)piperidin-1-yl)pyridin-3-yl)picolinamide

Following Method 11 of Example 305,tert-butyl(R)-4-(3-(tert-butyldimethylsilyloxy)piperidin-1-yl)pyridin-3-aminewas coupled to 3-amino-6-bromopicolinic acid yielding(R)-3-amino-6-bromo-N-(4-(3-(tert-butyldimethyl-silyloxy)piperidin-1-yl)pyridin-3-yl)picolinamide.LCMS (m/z): 506.2 (MH⁺); LC R_(t)=4.03 min.

Synthesis of benzyl 3-hydroxy-3-methylpiperidine-1-carboxylate

To a solution of benzyl 3-oxopiperidine-1-carboxylate (2.33 g, 10 mmol)in dry THF (50 mL) at −78° C. was added MeMgBr (3.6 mL, 3M solution inTHF, 11 mmol) slowly. The reaction was allowed to stir at −78° C. for 10min then slowly warmed to r.t. The reaction was quenched with NH₄Cl anddissolved in EtOAc (300 mL) and washed with saturated NH₄Cl and brine,dried and filtered, and concentrated to give the crude. The crudematerial was further purified by silica gel chromatography to yieldbenzyl 3-hydroxy-3-methylpiperidine-1-carboxylate. (53% yield). LCMS(m/z): 250.1 (MH⁺); LC R_(t)=2.98 min.

Synthesis of 3-methylpiperidin-3-ol

Method 17 was followed using benzyl3-hydroxy-3-methylpiperidine-1-carboxylate yielding3-methylpiperidin-3-ol (70%). LCMS (m/z): 116.1 (MH⁺).

Synthesis of 3-methyl-1-(3-nitropyridin-4-yl)piperidin-3-ol

Method 1 of Example 1 was followed using 1 equivalent each of4-chloro-3-nitropyridine, 3-methylpiperidin-3-ol and triethyl amine inDMF yielding 3-methyl-1-(3-nitropyridin-4-yl)piperidin-3-ol. LCMS (m/z):238.1 (MH⁺); LC R_(t)=1.39 min.

Synthesis of 1-(3-aminopyridin-4-yl)-3-methylpiperidin-3-ol

Following method 2 of Example 49,3-methyl-1-(3-nitropyridin-4-yl)piperidin-3-olin ethanol was reducedyielding 1-(3-aminopyridin-4-yl)-3-methylpiperidin-3-ol, (80%). LCMS(m/z): 208.1 (MH⁺); LC R_(t)=1.32 min.

Synthesis of Methyl 3-amino-5-fluoropicolinate

A solution of 2-bromo-5-fluoropyridin-3-amine (1.0 equiv.),triethylamine (1.6 equiv.), and Pd(BINAP)Cl₂ (0.0015 equiv.) inanhydrous methanol (0.4 M solution) in a sealed steel bomb was heated to100° C. After 3 h, more Pd catalyst (0.0015 equiv.) was added, thereaction mixture was re-heated to the same temperature for 3 h. Aftercooled down to room temperature, a brown precipitate was filtered offand the filtrate was extracted with EtOAc, which was washed with waterand brine, dried over anhydrous sodium sulfate, and filtered. Afterremoving volatile materials, the crude yellow product was obtained andused for the next step without further purification (40%). LCMS (m/z):271.2 (MH⁺); LC R_(t)=3.56 min.

Synthesis of Methyl 3-amino-6-bromo-5-fluoropicolinate

To a solution of methyl 3-amino-5-fluoropicolinate (1.0 equiv.) inacetonitrile (0.3 M solution) was added NBS (1.1 equiv.) for 2 minutesat room temperature. After quenched with water, the reaction mixture wasextracted with EtOAc. The crude product was purified by silica columnchromatography (20% to 50% EtOAc in hexanes) to give methyl3-amino-6-bromo-5-fluoropicolinate, (41%). LCMS (m/z): 249.1 (MH⁺); LCR_(t)=2.80 min.

Synthesis of 3-Amino-6-bromo-5-fluoropicolinic acid

To a solution of methyl 3-amino-5-fluoropicolinate (1.0 equiv.) intetra-hydrofuran and methanol (2:1, 0.2 M solution) was added LiOH (1.8equiv., 1 M aqueous solution) at room temperature. The reaction mixturewas stirred for 3 h and neutralized with 1.0 N aqueous HCl solution.Then, the reaction mixture was extracted with EtOAc, which was washedwith water and brine, dried over anhydrous sodium sulfate, and filtered.After removing volatile materials, the crude3-amino-6-bromo-5-fluoropicolinic acid was obtained and used for thenext step without further purification (92%). LCMS (m/z): 234.2 (MH⁺);LC R_(t)=2.25 min.

Synthesis of 3-amino-6-(2,6-difluorophenyl)-5-fluoropicolinic acid

To a solution of methyl 3-amino-5-fluoropicolinate (1.0 equiv.) inDME/2M Na₂CO₃ (3:1, 0.05 M) equipped with microwave vial was added2,6-difluorophenylboronic acid (3.0 equiv.) and Pd(dppf)Cl₂-DCM (0.1equiv.). The reaction mixture was heated to 140° C. for 10 min inmicrowave reactor. After 2,6-difluorophenylboronic acid (3.0 equiv.) wasadded more, the reaction mixture was heated once more to 140° C. for 10min in microwave reactor. After the reaction mixture was cooled to roomtemperature, H₂O and EtOAc were added and the organic phase was washedwith brine, then dried with Na₂SO₄, and concentrated. The crude materialwas purified via preparative HPLC. The pure methyl3-amino-6-(2,6-difluorophenyl)-5-fluoropicolinate was obtained after thepure fractions were neutralized with NaHCO₃, extracted with EtOAc, andconcentrated (34%). The methyl3-amino-6-(2,6-difluorophenyl)-5-fluoropicolinate (1.0 equiv.) wasdissolved in THF and MeOH

(2:1, 0.2 M) followed by addition of LiOH (1.8 equiv., 1 M aqueoussolution). After the reaction mixture was stirred for 1.5 h at roomtemperature, the reaction mixture was quenched with 1 N HCl solution(1.8 equiv.) and extracted with EtOAc. The organic phase was washed withbrine, then dried with Na₂SO₄, and concentrated. The crude3-amino-6-(2,6-difluorophenyl)-5-fluoropicolinic acid was used for thenext step without further purification (88%). LCMS (m/z): 269.0 (MH⁺);LC R_(t)=3.26 min.

Synthesis of3-amino-N-(4-chloropyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide

To a solution of 4-chloropyridin-3-amine (1.0 equiv.) and3-amino-6-(2,6-difluorophenyl)-5-fluoropicolinic acid (1.0 equiv.) inNMP (1 M) was added HOAt and EDCl sequentially. The reaction mixture wasstirred at room temperature for 2 days. The crude reaction was purifiedby prep HPLC to give3-amino-N-(4-chloropyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide(14%). LCMS (m/z): 379.0 (MH⁺); LC R_(t)=3.49 min

Synthesis of trans-(+/−)-tert-butyl1-(3-(3-amino-6-bromopicolinamido)pyridin-4-yl)-4-fluoropiperidin-3-ylcarbamate

Following Method 11 of Example 305, trans-(+/−)-tert-butyl1-(3-aminopyridin-4-yl)-4-fluoropiperidin-3-ylcarbamate and3-amino-6-bromopicolinic acid were reacted yielding after purification(+/−)-tert-butyl1-(3-(3-amino-6-bromopicolinamido)pyridin-4-yl)-4-fluoropiperidin-3-ylcarbamate,(20%). LCMS (m/z): 510.9 (MH⁺).

Synthesis of trans-(+/−)-tert-butyl1-(3-(3-amino-6-bromopicolinamido)pyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate

Following Method 11 of Example 305, trans-(+/−)-tert-butyl1-(3-aminopyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamateand 3-amino-6-bromo-picolinic acid was reacted yieldingtrans-(+/−)-tert-butyl1-(3-(3-amino-6-bromo-picolinamido)pyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate,(27%). LCMS (m/z): 621.2 (MH⁺); LC R_(t)=4.41 min.

Synthesis of (+/−)-tert-Butyl1-(3-(3-amino-6-bromopicolinamido)pyridin-4-yl)-3-(tert-butyldimethylsilyloxy)piperidin-4-ylcarbamate

Following Method 11 of Example 305, trans-(+/−)-tert-butyl1-(3-aminopyridin-4-yl)-3-(tert-butyldimethylsilyloxy)piperidin-4-ylcarbamateand 3-amino-6-bromo-picolinic acid was reacted yieldingtrans-(+/−)-tert-butyl1-(3-(3-amino-6-bromopicolinamido)pyridin-4-yl)-3-(tert-butyldimethylsilyloxy)piperidin-4-yl-carbamate,(20%). LCMS (m/z): 623.2 (MH⁺); LC R_(t)=4.12 min.

Synthesis of trans-tert-Butyl1-(3-(3-amino-6-bromo-5-fluoropicolinamido)pyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate

Following Method 11 of Example 305, trans-(+/−)-tert-butyl1-(3-aminopyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamateand 3-amino-6-bromo-5-fluoropicolinic acid was reacted yieldingtrans-(+/−)-tert-butyl1-(3-(3-amino-6-bromo-5-fluoropicolinamido)pyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-yl-carbamate.LCMS (m/z): 641.2 (MH⁺); LC R_(t)=4.47 min.

Synthesis of trans-(+/−)-tert-Butyl1-(3-(3-amino-6-bromo-5-fluoropicolinamido)pyridin-4-yl)-3-(tert-butyldimethylsilyloxy)piperidin-4-ylcarbamate

Following Method 11 of Example 305, trans-(+/−)-tert-butyl1-(3-aminopyridin-4-yl)-3-(tert-butyldimethylsilyloxy)piperidin-4-ylcarbamateand 3-amino-6-bromo-5-fluoropicolinic acid was reacted yieldingtrans-(+/−)-tert-butyl1-(3-(3-amino-6-bromo-5-fluoropicolinamido)pyridin-4-yl)-3-(tert-butyldimethylsilyloxy)piperidin-4-yl-carbamate.LCMS (m/z): 641.2 (MH⁺); LC R_(t)=4.73 min.

Method 20 Synthesis of5-amino-2-(2,6-difluorophenyl)pyrimidine-4-carboxylic acid

A 2.68 M NaOEt in EtOH solution (3 eq) was added to an ice-bath cooledmixture of 2,6-difluorobenzimidamide hydrochloride (2 eq) in EtOH (0.1M). The resulting mixture was allowed to warm to rt and stirred under N₂for 30 min. To the reaction mixture was added drop wise a solution ofmucobromic acid (1 eq) in EtOH and the reaction was heated in a 50° C.oil bath for 2.5 hr. After cooling to rt the reaction mixture wasconcentrated in vacuo. H₂O and 1.0 N NaOH were added and the aqueousmixture was washed with EtOAc. The aqueous phase was acidified to pH=4with 1.0 N HCl then extracted with EtOAc. Combined organic extracts werewashed once with brine, then dried over anhydrous Na₂SO₄, filtered, andconcentrated in vacuo to give5-bromo-2-(2,6-difluorophenyl)pyrimidine-4-carboxylic acid. The crudeproduct was used for the next step without further purification. LC/MS(m/z): 316.9 (MH⁺). LC: R_(t): 2.426 min.

CuSO₄ (0.1 eq) was added to a mixture of5-bromo-2-(2,6-difluorophenyl)pyrimidine-4-carboxylic acid (1 eq) and28% aqueous ammonium hydroxide solution in a microwave reaction vessel.The reaction mixture was heated in a microwave reactor at 110° C. for 25min. The reaction vessel was cooled in dry ice for 30 min then unsealedand concentrated in vacuo. To the resulting solids was added 1.0 N HCland the mixture was extracted with EtOAc. Combined organic extracts werewashed once with brine, then dried over anhydrous Na₂SO₄, filtered, andconcentrated in vacuo to give5-amino-2-(2,6-difluorophenyl)pyrimidine-4-carboxylic acid. The crudeproduct was used for the next step without further purification. LC/MS(m/z): 252.0 (MH⁺). LC: R_(t): 2.043 min.

Synthesis of 5-amino-2-(2-fluorophenyl)pyrimidine-4-carboxylic acid

Following method 20, 5-amino-2-(2-fluorophenyl)pyrimidine-4-carboxylicacid was prepared starting from 2-fluorobenzimidamide hydrochloride.LC/MS (m/z): 234.0 (MH⁺), R_(t): 0.70 min.

Synthesis of 5-amino-2-phenylpyrimidine-4-carboxylic acid

Following method 20, 5-amino-2-phenylpyrimidine-4-carboxylic acid wasprepared starting from benzimidamide hydrochloride. LC/MS (m/z): 216.1(MH⁺).

Synthesis of ethyl 5-amino-2-chloropyrimidine-4-carboxylate

10% Palladium on carbon (0.2 eq) was added to a N₂-flushed mixture ofethyl 2,6-dichloro-5-nitropyrimidine-4-carboxylate (1 eq) and magnesiumoxide (2 eq) in 1,4-dioxane (0.15 M). The reaction was purged with H₂under atmospheric pressure at rt. After 16 h additional portions of 10%Pd/C (0.3 eq) and MgO (5 eq) were added and the reaction continued topurge with H₂ under atmospheric pressure for 6 h at rt. The crude solidswere filtered through a pad of Celite on a paper lined Buchner funneland washed with CH₂Cl₂. The filtrate was transferred to a separatoryfunnel, washed twice with H₂O and once with brine, then dried overanhydrous Na₂SO₄, filtered, and concentrated in vacuo. The crude productwas dissolved in CH₂Cl₂, loaded onto aSiO_(2 column, and purified by flash chromatography ()10-20-30% EtOAc inhexanes) to give ethyl 5-amino-2-chloropyrimidine-4-carboxylate. LC/MS(m/z): 202.0 (MH⁺).

Synthesis of 5-amino-2-chloropyrimidine-4-carboxylic acid

A 0.5 M aqueous solution of LiOH (1.5 eq) was added to a stirringmixture of ethyl 5-amino-2-chloropyrimidine-4-carboxylate (1 eq) in H₂O(0.1 M) and THF (0.1M). The reaction was maintained for 2 h at rt. 1.0 NHCl was added and the crude mixture was concentrated in vacuo to removeresidual THF. The resulting solids were collected on a paper linedBuchner funnel and dried for 16 h under vacuum to give5-amino-2-chloropyrimidine-4-carboxylic acid. LC/MS (m/z): 174.0 (MH⁺).HPLC: R_(t): 1.148 min.

Synthesis of 3-nitro-5-phenylpicolinonitrile

5-bromo-3-nitropicolinonitrile (1 eq) and phenylboronic acid (1.5 eq)was mixed with 15 mL of 1,4-dioxane and 5 mL of 2 M Na₂CO₃ aqueoussolution in a glass pressure tube. The reaction mixture was degassed byanhydrous N₂ stream for 5 min followed by the addition ofPd(dppf)Cl₂-DCM (0.1 eq). The reaction mixture was stirred at 100° C.for 3 hours. Then the mixture was diluted with 100 mL of ethyl acetateand washed with water, brine, then dried over MgSO₄, filtered, andevaporated under reduced pressure to give crude product, which wastriturated by DCM, ether, hexanes to give the pure titled compound.LC/MS (m/z): 226.1 (MH⁺).

Synthesis of 3-amino-5-phenylpicolinic acid

To a solution of 3-nitro-5-phenylpicolinonitrile (1 eq) in 10 mL of DMFwas added tin (II) chloride dehydrate (7.0 eq) at room temperature. Thereaction mixture was stirred at that temperature overnight. The mixturewas diluted with 150 mL of ethyl acetate and 30 mL of triethyl amine.After filtration, the filtrate was concentrated under reduced pressureto give a solid, which was added 2 mL of concentrated HCl. The mixturewas stirred in microwave at 90° C. for 10 minutes. After standing overnight, the solid was collected by filtration, which was dissolved in 10mL of 1N NaOH. The resulting mixture was extracted with 50 mL of ethylacetate. The aqueous layer was acidified by 1 N HCl to pH 7.0 to yield3-amino-5-phenylpicolinic acid, which was collected by filtration. LC/MS(m/z): 215.1 (MH⁺).

Synthesis of 6-bromo-5-fluoropicolinic acid

To 2-bromo-3-fluoro-6-methylpyridine (2.0 g, 10.58 mmoles) in H₂O (30mL) was added potassium permanganate (1.67 g, 10.58 mmoles). Thesolution was heated at 100° C. for 5 hours at which time more potassiumpermanganate (1.67 g, 10.58 mmoles) was added. After heating for anadditional 48 hours the material was filtered through celite (4 cm×2inches) and rinsed with H₂O (150 mL). The combined aqueous was acidifiedwith 1N HCl to pH4, extracted with ethyl acetate (200 mL), washed withNaCl(sat.), dried over MgSO₄, filtered and concentrated to yield6-bromo-5-fluoropicolinic acid (17%) as a white solid.

LCMS (m/z): 221.9 (MH⁺); LC R_(t)=2.05 min.

Synthesis of (S)-tert-butyl1-(3-(6-bromo-5-fluoropicolinamido)pyridin-4-yl)piperidin-3-ylcarbamate

Following Method 11 (Example 305), (S)-tert-butyl1-(3-aminopyridin-4-yl)piperidin-3-ylcarbamate was coupled to6-bromo-5-fluoropicolinic acid yielding crude (S)-tert-butyl1-(3-(6-bromo-5-fluoropicolinamido)pyridin-4-yl)piperidin-3-ylcarbamate(92%) which was used as is. LCMS (m/z): 496.2 (MH⁺); LC Rt=2.90 min.

Synthesis of 6-bromo-3-fluoropicolinic acid

To 6-bromo-3-fluoro-2-methylpyridine (2.0 g, 10.58 mmoles) in H₂O (200mL) was added potassium permanganate (1.67 g, 10.58 mmoles). Thesolution was heated at 100° C. for 16 hours at which time upon coolingthe material was filtered through celite (4 cm×2 inches) and rinsed withH₂O (150 mL). The combined aqueous was acidified with 1N HCl to pH4,extracted with ethyl acetate (2×200 mL), washed with NaCl(sat.), driedover MgSO₄, filtered and concentrated to yield 6-bromo-3-fluoropicolinicacid (18%) as a white solid. LCMS (m/z): 221.9 (MH⁺); LC R_(t)=1.71 min.

Synthesis of (S)-tert-butyl1-(3-(6-bromo-3-fluoropicolinamido)pyridin-4-yl)piperidin-3-ylcarbamate

Following Method 11 (Example 305), (S)-tert-butyl1-(3-aminopyridin-4-yl)piperidin-3-ylcarbamate was coupled to6-bromo-3-fluoropicolinic acid yielding (S)-tert-butyl1-(3-(6-bromo-3-fluoropicolinamido)pyridin-4-yl)piperidin-3-ylcarbamatewhich was used directly as is. LCMS (m/z): 496.2 (MH⁺); LC R_(t)=2.71min.

Synthesis of (S)-tert-butyl1-(3-(6-bromopicolinamido)pyridin-4-yl)piperidin-3-ylcarbamate

Following Method 11 (Example 305), (S)-tert-butyl1-(3-aminopyridin-4-yl)piperidin-3-ylcarbamate was coupled to6-bromopicolinic acid yielding, after column chromatography (EtOAc aseluant), (S)-tert-butyl1-(3-(6-bromopicolinamido)pyridin-4-yl)piperidin-3-ylcarbamate (82%).LCMS (m/z): 478.1 (MH⁺); LC R_(t)=2.84 min.

Synthesis of (S)-tert-butyl1-(3-(5-amino-2-chloropyrimidine-4-carboxamido)pyridin-4-yl)piperidin-3-ylcarbamate

Following Method 11 (Example 305), (S)-tert-butyl1-(3-aminopyridin-4-yl)piperidin-3-ylcarbamate was coupled to5-amino-2-chloropyrimidine-4-carboxylic acid yielding, after columnchromatography (EtOAc as eluant), (S)-tert-butyl1-(3-(5-amino-2-chloropyrimidine-4-carboxamido)pyridin-4-yl)piperidin-3-yl-carbamate(10%). LCMS (m/z): 433.1 (MH⁺); LC R_(t)=2.46 min.

Method 21 Synthesis of 2-chloro-6-isobutoxypyrazine

A flame-dried round bottom flask was charged with a suspension of 95%NaH (1.1 eq) in anhydrous THF (0.3 M). The stirring mixture was cooledto 0° C. in an ice-water bath and 2-methyl-1-propanol (1 eq) was addeddrop wise via syringe. After 30 min 2,6-dichloropyrazine (1 eq) wasadded, the reaction was warmed to rt and stirred for 3 h. The crudemixture was quenched with saturated aqueous NH₄Cl and extracted withEtOAc. The combined organic phases were washed once each with H₂O andbrine, then dried over anhydrous Na₂SO₄, filtered, and concentrated invacuo. The crude residue was dissolved in CH₂Cl₂, loaded onto a SiO₂column, and purified by flash chromatography (9:1 hexanes/EtOAc eluent)to give 2-chloro-6-isobutoxypyrazine. LC/MS (m/z): 187.1 (MH⁺).

Synthesis of 2-chloro-6-(cyclopropylmethoxy)pyrazine

Following Method 21,2-chloro-6-(cyclopropylmethoxy)pyrazine wasprepared. LC/MS (m/z): 185.0 (MH⁺).

Synthesis of 2-chloro-6-ethoxypyrazine

Following Method 21,2-chloro-6-ethoxypyrazine was prepared. LC/MS (m/z):159.0 (MH⁺).

Synthesis of 2-chloro-6-isopropoxypyrazine

Following Method 21,2-chloro-6-isopropoxypyrazine was prepared. LC/MS(m/z): 173.1 (MH⁺).

Synthesis of 2-chloro-6-propoxypyrazine

Following Method 21,2-chloro-6-propoxypyrazine was prepared. LC/MS(m/z): 173.1 (MH⁺).

Synthesis of 2-(benzyloxy)-6-chloropyrazine

Following Method 21, 2-(benzyloxy)-6-chloropyrazine was prepared. LC/MS(m/z): 221.0 (MH⁺).

Method 22 Synthesis of 5-bromo-3-(2-methoxyethoxy)pyrazin-2-amine

A flame-dried round bottom flask was charged with a suspension of 95%NaH (1.3 eq) in anhydrous THF (0.2 M). The stirring mixture was cooledto 0° C. in an ice-water bath and 2-methoxyethanol (1.2 eq) was addeddrop wise via syringe. After 30 min 3,5-dibromopyrazin-2-amine (1 eq)was added, the reaction was warmed to rt and stirred for 3 h. The crudemixture was quenched with saturated aqueous NH₄Cl and extracted withEtOAc. The combined organic phases were washed once each with H₂O andbrine, then dried over anhydrous Na₂SO₄, filtered, and concentrated invacuo to give 5-bromo-3-(2-methoxyethoxy)pyrazin-2-amine LC/MS (m/z):250.0 (MH⁺).

Synthesis of 5-bromo-3-(2,2,2-trifluoroethoxy)pyrazin-2-amine

Following Method 22,5-bromo-3-(2,2,2-trifluoroethoxy)pyrazin-2-amine wasprepared. LC/MS (m/z): 274.0 (MH⁺).

Method 23 Synthesis of3-(2-methoxyethoxy)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazin-2-amine

To a solution of 5-bromo-3-(2-methoxyethoxy)pyrazin-2-amine (1 eq) indioxane (0.25 M) in a microwave reaction vessel was addedbispinacolatodiboron (2 eq), Pd(dba)₂ (0.05 eq), PCy₃ (0.075 eq) andKOAc (3 eq). The reaction mixture was then heated twice in a microwavereactor at 110° C. for 600 sec. The crude product was used for the nextstep without workup or further purification. LC/MS (m/z): 214.1/296.1(MH⁺).

Synthesis of5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-(2,2,2-trifluoroethoxy)pyrazin-2-amine

Following Method 23,5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-(2,2,2-trifluoroethoxy)pyrazin-2-aminewas prepared from 5-bromo-3-(2,2,2-trifluoroethoxy)pyrazin-2-amine LC/MS(m/z): 238.1 (MH⁺).

Method 24 Synthesis of tert-butyl6-chloro-2-methylpyrimidin-4-ylcarbamate

To a solution of 6-chloro-2-methylpyrimidin-4-amine (1.0 equiv.) in THF(0.17M) was added BOC₂O (1.1 equiv.) and DMAP (cat.). The reaction wasallowed to stir overnight, then concentrated to a yellow crude, andfiltered through a pad of SiO₂ eluting with EtOAc and hexanes (1:1) toafford an off-white solid (78%). LCMS (m/z): 244.1 (MH⁺); LC R_(t)=3.69min.

Synthesis of 4-chloro-N,N-di-BOC-6-methylpyrimidin-2-amine

Method 24 was followed using 4-chloro-6-methylpyrimidin-2-amine (1.0equiv.), BOC₂O (2.0 equiv.), and DMAP (cat.) to afford4-chloro-N,N-di-BOC-6-methylpyrimidin-2-amine in 71%. LCMS (m/z): 344.2(MH⁺); LC R_(t)=4.3 min.

Synthesis of 4-chloro-N,N-di-BOC-6-methoxypyrimidin-2-amine

Method 24 was followed using 4-chloro-6-methoxypyrimidin-2-amine (1.0equiv.), Boc₂O (2.0 equiv.), and DMAP (cat.) to afford4-chloro-N,N-di-BOC-6-methoxypyrimidin-2-amine in >95%. LCMS (m/z):360.2 (MH⁺); LC R_(t)=5.70 min.

Synthesis of 6-chloro-N,N-di-BOC-2-(methylthio)pyrimidin-4-amine

Method 24 was followed using 6-chloro-2-(methylthio)pyrimidin-4-amine(1.0 equiv.), BOC₂O (2.0 equiv.), and DMAP (cat.) to afford6-chloro-N,N-di-BOC-2-(methylthio)pyrimidin-4-amine in >95%. LCMS (m/z):376.1 (MH⁺); LC R_(t)=4.9 min.

Synthesis of tert-butyl6-chloro-2-(trifluoromethyl)pyrimidin-4-ylcarbamate

Method 24 was followed using6-chloro-2-(trifluoromethyl)pyrimidin-4-amine (1.0 equiv.), BOC₂O (1.0equiv.), and DMAP (cat.) to afford tert-butyl6-chloro-2-(trifluoromethyl)pyrimidin-4-ylcarbamate in 64%. LCMS (m/z):298.1 (MH⁺); LC R_(t)=4.73 min.

Synthesis of benzyl 4-chloropyridin-3-ylcarbamate

A solution of benzyl chloroformate (1.1 equiv.) in THF (1.85 M) wasslowly added to a solution of 3-amino-4-chloropyridine (1.0 equiv.) andpyridine (1.5 equiv.) in THF (1.0 M) and stirred at rt for 3.5 hours(formation of a precipitate over time). The reaction was quenched withH₂O (100 mL), extracted with EtOAc (200 mL), washed with NaCl_((sat))(75 mL), dried over MgSO₄, filtered and the volatiles were removed invacuo. The product precipitated from a mixture of hexane/EtOAc yieldingbenzyl 4-chloropyridin-3-ylcarbamate (34%). LCMS (m/z): 263.1 (MH⁺); LCR_(t)=2.33 min.

Synthesis of benzyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaboronlan.2-yl)pyridine-3-ylcarbamate

A solution of benzyl 4-chloropyridin-3-ylcarbamate (1.0 equiv.),bis(pinacolato)diboron (2.0 equiv.), Pd₂(dba)₃ (0.05 equiv.), PCy₃(0.075 equiv.), KOAc (2.0 equiv.) in dioxane (0.19 M) was degassed bybubbling nitrogen through for 10 min in a round-bottomed flask. Theflask was heated to 90° C. for 3 hours, cooled to room temperature,filtered through activated charcoal and Celite and washed with EtOAc.Upon concentration of the filtrate, a thick dark brown product wasobtained. LCMS (m/z): 273 (MH⁺ for the corresponding boronic acid); LCR_(t)=1.93 min.

Method 25 Synthesis of tert-butyl6-(3-(benzylcarbamate-amino)-pyridin-4-yl)-2-(trifluoromethyl)pyrimidin-4-ylcarbamate

To the crude solution of benzyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaboronlan.2-yl)pyridine-3-ylcarbamate(3.0 equiv.) was added Pd(dppf)Cl₂-DCM (0.10 equiv.), tert-butyl6-chloro-2-(trifluoromethyl)pyrimidin-4-ylcarbamate (1.0 equiv.), andDME/2M Na₂CO₃ (3:1, 0.08 M). The reaction was heated to 90° C. for onehour, then cooled to room temperature, H₂O and EtOAc were added, theorganic layer was extracted, washed with brine and dried with Na₂SO₄.Upon concentration, the crude was passed through a pad of SiO₂, washingwith EtOAc. The reaction was concentrated to almost dryness, and hexanewas added. The precipitate was filtered to give the product as a lightyellow powder. The filtrate was concentrated to almost dryness, addedmore hexane and filtered the precipitate. Total yield=50%. LCMS (m/z):490.1 (MH⁺); LC R_(t)=4.11 min.

Synthesis of benzyl4-(2-(di-BOC-amino)-6-methylpyrimidin-4-yl)pyridin-3-ylcarbamate

Method 25 was followed using4-chloro-N,N-di-BOC-6-methylpyrimidin-2-amine (1.0 equiv.), benzyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaboronlan.2-yl)pyridine-3-ylcarbamate(3 equiv.), Pd(dppf)Cl₂-DCM (0.10 equiv.) in DME/2M Na₂CO₃ (3:1, 0.07M)at 70° C. for 30 min. Purification via SiO₂ column chromatographyeluting with EtOAc and hexanes (2.5:1) afforded benzyl4-(2-(di-BOC-amino)-6-methylpyrimidin-4-yl)pyridin-3-ylcarbamate in 69%yield. LCMS (m/z): 536.2 (MH⁺); LC R_(t)=4.2 min.

Method 26 Synthesis of tert-butyl6-(3-aminopyridin-4-yl)-2-(trifluoromethyl)pyrimidin-4-ylcarbamate

A solution of tert-butyl6-(3-(benzylcarbamate-amino)pyridin-4-yl)-2-(trifluoromethyl)pyrimidin-4-ylcarbamatewas stirred in EtOAc and EtOH (3:1, M) (heterogeneous solution). Pd/C(10% by weight) was added and the reaction was stirred under a hydrogenballoon for 2 days. Upon completion, the solution was filtered throughCelite and washed with EtOAc. The filtrate was concentrated to give abrown solid (>95%). LCMS (m/z): 356.1 (MH⁺); LC R_(t)=2.80 min.

Synthesis of4-(3-aminopyridin-4-yl)-N,N-di-BOC-6-methylpyrimidin-2-amine

Method 26 was followed using benzyl4-(2-(di-BOC-amino)-6-methylpyrimidin-4-yl)pyridin-3-ylcarbamate (1.0equiv.), Pd/C (20% by weight) in EtOAc yielding4-(3-aminopyridin-4-yl)-N,N-di-BOC-6-methylpyrimidin-2-amine in 90%yield. LCMS (m/z): 402.3 (MH⁺); LC R_(t)=3.0 min.

Synthesis of4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-amine

A solution of benzyl 4-chloropyridin-3-ylcarbamate (1.0 equiv.),bis(pinacolato)diboron (2.0 equiv.), Pd₂(dba)₃ (0.05 equiv.), PCy₃(0.075 equiv.), KOAc (2 equiv.) in dioxane (0.19 M) was degassed bybubbling nitrogen through for 10 min in a round-bottomed flask. Theflask was heated to 90° C. for 16 hours, cooled to room temperature,filtered through activated charcoal and Celite, washed with EtOAc andconcentrated to give a thick dark brown product. LCMS (m/z): 139.0 (MH⁺for the corresponding boronic acid).

Synthesis of6-(3-aminopyridin-4-yl)-N,N-di-BOC-2-(methylthio)pyrimidin-4-amine

Method 26 was followed using6-chloro-N,N-di-BOC-2-(methyl-thio)pyrimidin-4-amine (1.0 equiv.),4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-amine (3.0equiv.), Pd(dppf)C12-DCM (0.10 equiv.) in DME/2M Na₂CO₃ (0.07M) at 90°C. for 30 min. Purification via SiO₂ column chromatography eluting withEtOAc and hexanes (1:1) afforded6-(3-aminopyridin-4-yl)-N,N-di-BOC-2-(methyl-thio)pyrimidin-4-amine in32% yield. LCMS (m/z): 434.2 (MH⁺); LC R_(t)=3.56 min.

Synthesis of tert-butyl6-(3-aminopyridin-4-yl)-2-methylpyrimidin-4-ylcarbamate

Method 26 was followed using tert-butyl6-chloro-2-methylpyrimidin-4-ylcarbamate (1.0 equiv.),4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-amine (3.0equiv.), Pd(dppf)Cl₂-DCM (0.10 equiv.) in DME/2M Na₂CO₃ (3:1, 0.07M) at80° C. for 30 min. Purification via SiO₂ column chromatography elutingwith EtOAc afforded tert-butyl6-(3-aminopyridin-4-yl)-2-methylpyrimidin-4-ylcarbamate in 26% yield.LCMS (m/z): 302.1 (MH⁺); LC R_(t)=2.23 min.

Synthesis of4-(3-aminopyridin-4-yl)-6-methoxy-N,N-di-BOC-pyrimidin-2-amine

Method 26 was followed using 4-chloro-N,N di BOC 6methoxypyrimidin-2-amine (1.0 equiv.),4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-pyridin-3-amine (3.0equiv.), Pd(dppf)Cl₂-DCM (0.10 equiv.) in DME/2M Na₂CO₃ (0.07M) at 90°C. for 30 min. Purification via SiO₂ column chromatography eluting withEtOAc and hexanes (1:1) afforded4-(3-aminopyridin-4-yl)-6-methoxy-N,N-di-BOC-pyrimidin-2-amine in 13%yield). LCMS (m/z): 418.1 (MH⁺).

Synthesis ofN²-(3,4-dimethoxybenzyl)-6-(trifluoromethyl)-4,4′-bipyridine-2,3′-diamine

Method 26 was followed usingN-(3,4-dimethoxybenzyl)-4-iodo-6-(tri-fluoromethyl)pyridin-2-amine (1.0equiv.), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-amine(3 equiv.), Pd(dppf)Cl₂DCM (0.10 equiv.) in DME/2M Na₂CO₃ (3:1, 0.07 M)at 50° C. for 45 min. Purification via reverse phase HPLC affordedN²-(3,4-dimethoxybenzyl)-6-(trifluoromethyl)-4,4′-bipyridine-2,3′-diaminein 38% yield. LCMS (m/z): 402.1 (MH⁺); LC R_(t)=3.0 min.

Method 27 Synthesis of tert-butyl6-(3-(3-amino-6-bromopicolinamido)pyridine-4-O-2-(trifluoromethyl)pyrimidin-4-ylcarbamate

A solution of tert-butyl6-(3-aminopyridin-4-yl)-2-(trifluoro-methyl)pyrimidin-4-ylcarbamate (1.0equiv.), 3-amino-6-bromopicolinic acid (1.0 equiv.), HOAt (1.0 equiv.),and EDC (1.0 equiv.) in DMF at a concentration of (0.2 M) was stirredfor 3 hrs then heated to 50° C. overnight (homogeneous solution). Waterwas added to the reaction and the precipitate was filtered. The solidwas further purified via SiO₂ column chromatography eluting withDCM/MeOH (10%) to yield a brown solid as the desired product (81%). LCMS(m/z): 554.1/556.1 (MH⁺); LC R_(t)=3.77 min.

Synthesis of3-amino-6-bromo-N-(4-(6-(di-BOC-amino)-2-(methylthio)pyrimidin-4-yl)pyridin-3-yl)picolinamide

Method 27 was followed using6-(3-aminopyridin-4-yl)-N,N-di-BOC-2-(methylthio)pyrimidin-4-amine (1.0equiv.), 3-amino-6-bromopicolinic acid (1.0 equiv), EDC (1.0 equiv.),and HOAt (1.0 equiv.) in DMF yielding3-amino-6-bromo-N-(4-(6-(di-BOC-amino)-2-(methylthio)pyrimidin-4-yl)pyridin-3-yl)picolinamidein 30% yield. LCMS (m/z): 632.1/634.0 (MH⁺); LC R_(t)=4.55 min.

Synthesis of tert-butyl6-(3-(3-amino-6-bromopicolinamido)pyridine-4-yl)-2-methylpyrimidin-4-ylcarbamate

Method 27 was followed using tert-butyl6-(3-aminopyridin-4-yl)-2-methylpyrimidin-4-ylcarbamate (1.0 equiv.),3-amino-6-bromopicolinic acid (1.0 equiv), EDC (1.0 equiv.), and HOAt(1.0 equiv.) in DMF yielding tert-butyl64343-amino-6-bromopicolinamido)pyridine-4-yl)-2-methylpyrimidin-4-ylcarbamatein 74% yield. LCMS (m/z): 499.9/501.9 (MH⁺); LC R_(t)=3.36 min.

Synthesis of3-amino-6-bromo-N-(4-(2-(di-BOC-amino)-6-methylpyrimidin-4-yl)pyridin-3-yl)picolinamide

Method 27 was followed using4-(3-aminopyridin-4-yl)-N,N-di-BOC-6-methylpyrimidin-2-amine (1.0equiv.), 3-amino-6-bromopicolinic acid (1.0 equiv), EDC (1.0 equiv.),and HOAt (1.0 equiv.) in DMF yielding3-amino-6-bromo-N-(4-(2-(di-BOC-amino)-6-methylpyrimidin-4-yl)pyridin-3-yl)picolinamidein 12% yield. LCMS (m/z): 602.2 (MH⁺); LC R_(t)=3.60 min.

Synthesis of tert-butyl6-(3-(3-amino-6-bromo-5-fluoropicolinamido)pyridin-4-yl)-2-methylpyrimidin-4-ylcarbamate

Method 27 was followed using tert-butyl6-(3-aminopyridin-4-yl)-2-methylpyrimidin-4-ylcarbamate (1.0 equiv.),3-amino-6-bromo-5-fluoropicolinic acid (1.0 equiv.), EDC (1.0 equiv.),and HOAt (1.0 equiv.) in DMF yielding tert-butyl64343-amino-6-bromo-5-fluoropicolinamido)pyridin-4-yl)-2-methylpyrimidin-4-ylcarbamatein 15% yield. LCMS (m/z): 520.1 (MH⁺); LC R_(t)=3.4 min.

Synthesis of tert-butyl4-(3-(3-amino-6-bromo-5-fluoropicolinamido)pyridin-4-yl)-6-methylpyrimidin-2-ylcarbamate

Method 27 was followed using tert-butyl4-(3-aminopyridin-4-yl)-2-methylpyrimidin-4-ylcarbamate (1.0 equiv.),3-amino-6-bromo-5-fluoropicolinic acid (1.0 equiv.), EDC (1.0 equiv.),and HOAt (1.0 equiv.) in DMF yielding tert-butyl44343-amino-6-bromo-5-fluoropicolinamido)pyridin-4-yl)-6-methylpyrimidin-2-ylcarbamate20% yield. LCMS (m/z): 618.1 (MH⁺); LC R_(t)=3.5 min.

Synthesis of tert-butyl6-(3-(6-bromopicolinamido)pyridin-4-yl)-2-(trifluoromethyl)pyrimidin-4-ylcarbamate

Method 27 was followed using tert-butyl6-(3-aminopyridin-4-yl)-2-(trifluoromethyl)pyrimidin-4-ylcarbamate (1.0equiv.), 6-bromopicolinic acid (1.0 equiv.), EDC (1.0 equiv.), and HOAt(1.0 equiv.) in NMP yielding tert-butyl64346-bromopicolinamido)pyridin-4-yl)-2-(trifluoromethyl)pyrimidin-4-ylcarbamate >95%yield. LCMS (m/z): 539/541 (MH⁺); LC R_(t)=3.97 min.

Synthesis of 2-chloro-3-iodo-6-(trifluoromethyl)pyridine

To a cooled solution (−78° C.) of n-BuLi (1.0 equiv.) in THF (0.8M) wasadded diisopropylamine (1.0 equiv.) dropwise, maintaining the internaltemperature under −70 oC. The solution was stirred for 30 min, then2-chloro-6-(trifluoromethyl)pyridine (1.0 equiv.) in THF was addeddropwise. The solution was stirred for another 30 min, then 12 was addedas a solid. Stirred for 1 hr at −78° C., then allowed the reaction towarm to room temperature overnight. The solution was quenched byaddition of H₂O, extracted with EtOAc, then washed with brine, andconcentrated. The crude material was purified via SiO₂ columnchromatography eluting with EtOAc and hexanes (1:10) to yield2-chloro-3-iodo-6-(trifluoromethyl)pyridine in 35% yield. LCMS (m/z):307.8 (MH⁺); LC R_(t)=4.18 min.

Synthesis of 2-chloro-4-iodo-6-(trifluoromethyl)pyridine

To a cooled (−75° C.) solution of THF was added n-BuLi (1.1 equiv.),followed by diisopropylamine (1.1 equiv.), dropwise. The reaction wasstirred for 10 min, then 2-chloro-3-iodo-6-(trifluoromethyl)pyridine(1.0 equiv.) was added dropwise in THF. The solution was stirred at −75°C. for 1 hr, then quenched with the addition of 1N HCl, extracted withEtOAc, washed with brine, dried with MgSO₄, and concentrated to give atan solid in 85% yield. LCMS (m/z): 307.8 (MH⁺); LC R_(t)=4.28 min.

Synthesis ofN-(3,4-dimethoxybenzyl)-4-iodo-6-(trifluoromethyl)pyridin-2-amine

A solution of 2-chloro-4-iodo-6-(trifluoromethyl)pyridine (1.0 equiv.),(3,4-dimethoxyphenyl)methanamide (5.0 equiv.), and Et₃N (5.0 equiv.) inNMP (0.7M) was microwaved to 100° C. for 10 min. The solution wasdirectly purified via reverse phase HPLC, the pure fractions wereneutralized with solid NaHCO₃, extracted with EtOAc, dried with MgSO₄,and concentrated to giveN-(3,4-dimethoxybenzyl)-4-iodo-6-(trifluoromethyl)pyridin-2-amine in 36%yield. LCMS (m/z): 347.1 (MH⁺); LC R_(t)=3.96 min.

Synthesis of methyl 3-amino-6-(2,6-difluorophenyl)picolinate

A solution of methyl 3-amino-6-bromopicolinate (1.0 equiv.),2,6-difluorophenyl-boronic acid (3.0 equiv), and Pd(dppf)Cl₂-DCM (0.1equiv.) in 3:1 DME/2M Na₂CO₃ (0.5M) was subjected to microwaveirradiation at 120° C. for 15 min intervals. The reaction was filteredand washed with EtOAc. The organic was partitioned with H₂O (25 mL), wasfurther washed with NaCl_((sat)) (25 mL), was dried over MgSO₄, and thevolatiles were removed in vacuo. The residue was diluted in EtOAc andpassed through a silica gel plug and the volatiles were removed in vacuoyielding methyl 3-amino-6-(2,6-difluorophenyl)picolinate (47%). LCMS(m/z): 265.1 (MH⁺); LC R_(t)=2.70 min.

Synthesis of 3-amino-6-(2,6-difluorophenyl)picolinic acid

To a solution of methyl 3-amino-6-(2,6-difluorophenyl)picolinate (1.0equiv) in THF (0.5M), was added 1M LiOH (4.0 equiv). After stirring for4 hours at 60° C., 1 N HCl (4.0 equiv.) was added and the THF wasremoved in vacuo. The resulting solid was filtered and rinsed with coldH₂O (3×20 mL) to yield 3-amino-6-(2,6-difluorophenyl)picolinic acid(90%). LCMS (m/z): 251.1 (MH⁺); LC R_(t)=2.1 min.

Synthesis of methyl 3-amino-6-(thiazol-2-yl)picolinate

A solution of methyl 3-amino-6-bromopicolinate (1.0 equiv.),2-thiazolylzinc bromide 0.5 M solution in THF (3.0 equiv.), andPd(dppf)Cl₂-DCM (0.05 equiv.) was stirred at 80° C. for 1.5 hours. Thereaction was filtered and washed with EtOAc. The organic was washed withH₂O (100 mL), and further washed with NaCl_((sat)) (50 mL), dried overMgSO₄, and the volatiles were removed in vacuo. The product wascrystallized with hexane/EtOAc (1:1) to yield methyl3-amino-6-(thiazol-2-yl)picolinate (51%). LCMS (m/z): 236.1 (MH⁺); LCR_(t)=2.3 min.

Synthesis of 3-amino-6-(thiazol-2-yl)picolinic acid

To a solution of methyl 3-amino-6-(thiazol-2-yl)picolinate (1.0 equiv)in THF (0.5M), was added 1M LiOH (4.0 equiv). After stirring for 4 hoursat 60° C., 1 N HCl (4.0 equiv.) was added and the THF was removed invacuo. The resulting solid was filtered and rinsed with cold H2O (3×20mL) to yield 3-amino-6-(thiazol-2-yl)picolinic acid (61%). LCMS (m/z):222.1 (MH⁺); LC Rt=1.9 min.

Synthesis of methyl3-amino-6-(2-fluoro-5-isopropylcabamoyl)phenyl)picolinate

A solution of methyl 3-amino-6-bromopicolinate (1.0 equiv.),N-iso-propyl 3-borono-4-fluorobenzamide (1.1 equiv.), andPd(dppf)Cl₂-DCM (0.15 equiv.) in DME/2M Na₂CO₃ (3:1), at a concentrationof 0.5 M, was stirred at 120° C. for 1.5 hours.

The reaction was filtered and washed with EtOAc. The organic waspartitioned with H₂O (25 mL), washed with NaCl_((sat)) (25 mL), driedover MgSO₄, and the volatiles were removed in vacuo. The residue wasdiluted in EtOAc and passed through a silica gel plug and the volatileswere removed in vacuo yielding methyl3-amino-6-(2-fluoro-5-iso-propylcabamoyl)phenyl)picolinate (60%). LCMS(m/z): 332.2 (MH⁺); LC R_(t)=2.9 min.

Synthesis of 3-amino-6-(2-fluoro-5-isopropylcabamoyl)phenyl)picolinicacid

To a solution of methyl3-amino-6-(2-fluoro-5-isopropylcabamoyl)phenyl)picolinate (1.0 equiv) inTHF (0.5M), was added 1M LiOH (4.0 equiv). After stirring for 4 hours at60° C., 1 N HCl (4.0 equiv.) was added and the THF was removed in vacuo.The resulting solid was filtered and rinsed with cold H₂O (3×20 mL) toyield 3-amino-6-(2-fluoro-5-isopropylcabamoyl)phenyl)picolinic acid(98%). LCMS (m/z): 318.1 (MH⁺); LC R_(t)=2 4 min

Synthesis of3-amino-N-(4-chloropyridin-3-yl)-6-(2,6-difluorophenyl)picolinamide

A solution of 3-amino-6-(2,6-difluorophenyl)picolinic acid (1.0 equiv.),4-chloropyridin-3-amine (2.0 equiv.), HOAt (1.0 equiv.), and EDC (1.0equiv.) in DCM at a concentration of (0.2M) was stirred for 24 hr. Waterwas added to the reaction, followed by EtOAc. The organic layer wasseparated, dried with brine, MgSO₄, and concentrated. The crude materialwas purified via SiO₂ column chromatography eluting with EtOAc andhexanes (1:1) to give the product as a light yellow solid (21% yield).LCMS (m/z): 361.1 (MH⁺); LC R_(t)=3.28 min.

Method 28 Synthesis of3-amino-6-(2,6-difluorophenyl)-N-(3′-fluoro-4,4′-bipyridin-3-yl)picolinamide

A solution of3-amino-N-(4-chloropyridin-3-yl)-6-(2,6-difluoro-phenyl)picolinamide(1.0 equiv.), 3-fluoropyridin-4-yl boronic acid (3.0 equiv.), andPd(dppf)Cl₂-DCM (0.10 equiv.) in DME/2M Na₂CO₃ (3:1) was heated to 120°C. with microwave irradiation for 10 min. Upon cooling, the reaction wasextracted with EtOAc, the organic layer was dried with Na₂SO₄ andconcentrated. The crude material was purified via reverse phase HPLC.The product fraction was lyophilized to give3-amino-6-(2,6-difluorophenyl)-N-(3′-fluoro-4,4′-bipyridin-3-yl)picolinamideas the TFA salt in 12% yield. LCMS (m/z): 404.1 (MH⁺); LC R_(t)=2.92min.

The following compounds were prepared using Method 28:

Ex- ample Structure Name MH+ LC 601

3-amino-N-(5-fluoro- 3,4′-bipyridin-3′-yl)-6-(2-fluorophenyl)pyridine-2- carboxamide 404.1 2.96 602

3-amino-6-(2- fluorophenyl)-N-(4- pyrimidin-5-ylpyridin-3-yl)pyridine-2-carboxamide 387.1 2.62 603

3-amino-N-[6-amino-5- (tri-fluoromethyl)-3,4′- bipyridin-3′-yl]-6-(2-fluorophenyl)pyridine-2- carboxamide 469.1 2.90 604

3-amino-N-(6-amino-3,4′- bipyridin-3′-yl)-6-(2-fluoro-phenyl)pyridine-2- carboxamide 401.1 2.04 605

3-amino-N-(2′-amino-4,4′- bipyridin-3-yl)-6-(2- fluorophenyl)pyridine-2-carboxamide 401.1 2.13 606

3-amino-N-4,4′-bipyridin- 3-yl-6-(2-fluorophenyl)-pyridine-2-carboxamide 386.1 2.36 607

3-amino-N-(2′,6′-difluoro- 4,4′-bipyridin-3-yl)-6-(2,6-difluorophenyl)pyridine-2- carboxamide 440.1 3.34 608

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2- carboxylic acid (6′-amino-2′-methoxy-[4,4′]bi- pyridinyl-3-yl)-amide 449.1 2.44 609

3-amino-6-(2- fluorophenyl)-N-(5- methoxy-3,4′-bipyridin-3′-yl)pyridine-2-carboxamide 416.2 2.65 610

3-amino-6-(2- fluorophenyl)-N-(3′- methoxy-4,4′-bipyridin-3-yl)pyridine-2-carboxamide 416.1 2.47 611

3-amino-6-(2- fluorophenyl)-N-(2- methoxy-3,4′-bipyridin-3′-yl)pyridine-2-carboxamide 416.1 3.07 612

3-amino-6-(2,6- difluorophenyl)-N-(2′- hydroxy-5′-methyl-4,4′-bipyridin-3-yl)pyridine-2- carboxamide 434.0 2.22 613

3-amino-6-(2- fluorophenyl)-N-(3′- methyl-4,4′-bipyridin-3-yl)pyridine-2-carboxamide 400.1 2.31 614

3-amino-6-(2- fluorophenyl)-N-(2′- hydroxy-4,4′-bipyridin-3-yl)pyridine-2-carboxamide 402.0 2.22 615

3-amino-N-(3′-chloro-4,4′- bipyridin-3-yl)-6-(2-fluorophenyl)pyridine-2- carboxamide 420.1 3.02 616

3-amino-N-(3′-fluoro-4,4′- bipyridin-3-yl)-6-(2-fluorophenyl)pyridine-2- carboxamide 404.1 2.92 617

3-amino-N-(2′-amino-4,4′- bipyridin-3-yl)-6-(2,6-difluorophenyl)pyridine-2- carboxamide 419.1 2.09 618

3-amino-N-(2′-cyano-4,4′- bipyridin-3-yl)-6-(2,6-difluorophenyl)pyridine-2- carboxamide 429.1 2.93

Method 29 Synthesis of3-amino-N-(4-(6-aminopyrazin-2-yl)pyridine-3-yl)-6-(2-fluorophenyl)picolinamide

A solution of3-amino-N-(4-chloropyridin-3-yl)-6-(2,6-difluoro-phenyl)picolinamide(1.0 equiv.), bis(pinacolato)diboron (2.0 equiv.), Pd(dppf)Cl₂-DCM (0.10equiv.), KOAc (2.0 equiv.) in dioxane (0.19 M) was stirred in themicrowave for 5 min at 120° C. then 10 min at 120° C. The reaction wasfiltered and concentrated. To the crude was added6-chloropyrazin-2-amine (2.0 equiv.) and more Pd(dppf)Cl₂-DCM (0.10equiv.) in DME/2M Na₂CO₃ (3:1, 0.1M). The reaction was heated to 100° C.in the oil bath for 2 hrs. Cooled to room temperature, added H₂O andEtOAc, the organic layer was extracted, dried with brine and Na₂SO₄, andconcentrated. The crude mixture was purified via reverse-phase HPLC andthe pure fractions were lyophilized to give3-amino-N-(4-(6-aminopyrazin-2-yl)pyridine-3-yl)-6-(2-fluorophenyl)picolinamideas the TFA salt in 19% yield. LCMS (m/z): 402.1 (MH⁺); LC R_(t)=2.58min.

The following compounds were prepared using Method 29:

Ex- ample Structure Name MH+ LC 619

3-amino-N-(2′-amino-6′- methyl-4,4′-bipyridin-3-yl)-6-(2,6-difluorophenyl)-5- fluoropyridine-2- carboxamide 451.2 2.29 620

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2- carboxylic acid (6′-amino-2′-ethyl-[4,4′]bipyridinyl-3- yl)-amide 447.1 2.34 621

3-amino-N-[2′-chloro-6′- (trifluoromethyl)-4,4′- bipyridin-3-yl]-6-(2,6-difluorophenyl)pyridine-2- carboxamide 506.1 3.75 622

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2- carboxylic acid (6-amino-4-methoxy-[2,4′]bipyridinyl- 3′-yl)-amide 449.1 2.46 623

3-amino-N-(2′-amino-6′- methyl-4,4′-bipyridin-3-yl)-6-(2,6-difluorophenyl)pyridine- 2-carboxamide 433.1 2.28 624

3-amino-N-(2′-amino-6′- methyl-4,4′-bipyridin-3-yl)-6-(2-fluorophenyl)pyridine-2- carboxamide 2.0 2.22 625

3-amino-N-[4-(6-amino- pyrazin-2-yl)pyridin-3-yl]-6-(2-fluorophenyl)pyridine-2- carboxamide 402.1 2.58 626

3-amino-N-[4-(2-amino-6- methylpyrimidin-4-yl)-pyridin-3-yl]-6-(2-fluoro- phenyl)pyridine-2- carboxamide 416.1 2.62 627

3-amino-N-[4-(2,6- diaminopyrimidin-4- yl)pyridin-3-yl]-6-(2-fluorophenyl)pyridine-2- carboxamide 417.0 2.13 628

3-amino-N-[4-(2- aminopyrimidin-4-yl)pyridin- 3-yl]-6-(2,6-difluorophenyl)pyridine-2- carboxamide 420.1 2.51 629

3-amino-N-[4-(6- aminopyrimidin-4-yl)pyridin- 3-yl]-6-(2,6-difluoro-phenyl)pyridine-2- carboxamide 420.1 2.56 630

3-amino-N-(2-amino-3,4′- bipyridin-3′-yl)-6-(2,6-difluorophenyl)pyridine-2- carboxamide 419.1 2.12

Method 30 Synthesis of(s)-3-amino-N-(4-(3-aminopiperidin-1-yl)pyridine-3-yl)-6-cyclohexylpicolinamide

A solution of (S)-tert-butyl1-(3-(3-amino-6-bromo-picolinamido)pyridin-4-yl)piperidin-3-ylcarbamate(1.0 equiv.), cyclohexylzinc bromide (0.5 M solution in THF, 3.0 equiv),Pd₂(dba)₃ (0.1 equiv.), and P(2-furyl)₃ (0.2 equiv.) were heated to 65°C. for 18 hrs. Two more equivalents of zinc bromide reagent were addedif the reaction was not complete after 18 hrs. The mixture was cooled tort and concentrated to give the crude material. The crude mixture wasthen stirred in DCM/TFA (25%) until completion. Concentrated thereaction to dryness and purified via reverse phase HPLC. The purefractions were lyophilized to give the TFA salt product (40%). LCMS(m/z): 395.3 (MH⁺); LC R_(t)=2.34 min.

Synthesis of3-amino-N-(4-(6-amino-2-(trifluoromethyl)pyrimidin-4-yl)pyridin-3-yl)-6-(thiazol-2-yl)picolinamide

Following method 30, a solution of tert-butyl6-(3-(3-amino-6-bromopicolinamido)pyridine-4-yl)-2-(trifluoromethyl)pyrimidin-4-ylcarbamate(1.0 equiv.), 2-thiazolyl-zincbromide (3.5 equiv.), and Pd(dppf)Cl₂-DCM(0.10 equiv.) in THF was microwaved at 100° C. for 15 min. The reactionwas concentrated to dryness under vacuo, then stirred in DCM/TFA (25%)for two hours. Upon concentration and purification via reverse phaseHPLC,3-amino-N-(4-(6-amino-2-(trifluoro-methyl)pyrimidin-4-yl)pyridin-3-yl)-6-(thiazol-2-yl)picolinamidewas obtained as the TFA salt in 48% yield. LCMS (m/z): 459.1 (MH⁺); LCR_(t)=2.46 min.

The following compounds were also prepared using Method 30:

Ex- ample Structure Name MH+ LC 631

N-{4-[(3S)-3-amino- piperidin-1-yl]pyridin-3-yl}-6-(1,3-thiazol-2-yl)pyridine- 2-carboxamide 381.1 1.54 632

3-amino-N-{4-[(3S)-3- aminopiperidin-1-yl]pyridin-3-yl}-6-(1,3-thiazol-2-yl)- pyridine-2-carboxamide 396.1 1.56 633

3-Amino-6-cyclohexyl- pyridine-2-carboxylic acid[4-(6-amino-2-trifluoro- methyl-pyrimidin-4-yl)- pyridin-3-yl]-amide457.9 3.44 634

3-Amino-6-(1-ethyl-propyl)- pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)- amide 383.22.44 635

6-Cyclohexyl-pyridine-2- carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)- amide 380.2 2.24 636

3-Amino-6-cyclohexyl- pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)- amide 395.32.34 637

3-amino-N-{4-[(3S)-3- aminopiperidin-1-yl]pyridin-3-yl}-6-methylpyridine-2- carboxamide 327.1 1.38 638

3-amino-N-{4-[(3S)-3- aminopiperidin-1-yl]pyridin- 3-yl}-6-cyclopropyl-pyridine-2-carboxamide 353.1 1.70 639

3-amino-N-{4-[(3S)-3- aminopiperidin-1-yl]pyridin-3-yl}-6-ethylpyridine-2- carboxamide 341.1 1.59 640

3-amino-N-[2′-amino-6′- (trifluoromethyl)-4,4′- bipyridin-3-yl]-6-(1,3-thiazol-2-yl)pyridine-2- carboxamide 458.1 2.59 641

3-amino-N-(2′-amino-6′- methyl-4,4′-bipyridin-3-yl)-6-(1,3-thiazol-2-yl)pyridine- 2-carboxamide 404.1 1.79 642

3-amino-N-{4-[6-amino-2- (trifluoromethyl)pyrimidin-4-yl]pyridin-3-yl}-6-(1,3- thiazol-2-yl)pyridine-2- carboxamide 459.12.46 643

3-amino-N-[4-(6-amino-2- methylpyrimidin-4- yl)pyridin-3-yl]-6-(1,3-thiazol-2-yl)pyridine-2- carboxamide 405.0 1.74 644

3-Amino-6-cyclopentyl- pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)- amide 381.32.13 645

6-Adamantan-1-yl-3-amino- pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)- amide 447.22.70 646

3-Amino-6- bicyclo[2.2.1]hept-2-yl- pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)- amide 407.22.37 647

N-{4-[6-amino-2-(trifluoro- methyl)pyrimidin-4-yl]-pyridin-3-yl}-6-(1,3-thiazol- 2-yl)pyridine-2-carboxamide 444.1 2.67

Synthesis of 4-chloro-6-methylpyridin-2-amine

To a 10% aqueous solution of dioxane (0.1 M) was added4,6-dichloropyridin-2-amine (1.0 equiv.), trimethylboroxine (1.5equiv.), Pd(PPh₃)₄ (0.10 equiv.) and K₂CO₃ (3.0 equiv.). The solutionwas heated in an oil bath to 120° C. for 18 hrs, cooled to roomtemperature (not all of starting material was consumed), extracted withEtOAc, dried with Na₂SO₄, and concentrated. The crude material waspurified via SiO₂ column chromatography eluting with 5% MeOH/DCM toyield an off-white solid in 23% yield. LCMS (m/z): 143 (MH⁺); LCR_(t)=1.11 min.

Synthesis of 6-methyl-3′-nitro-4,4′-bipyridin-2-amine

To a solution of 4-chloro-6-methylpyridin-2-amine (1.0 equiv.) indioxane was added bis(pinacolato)diboron (2.0 equiv.), Pd₂(dba)₃ (0.05equiv.), PCy₃ (0.075 equiv.), and KOAc (3.0 equiv.). The reaction washeated for 3 hrs at 110° C., then filtered, and concentrated. The crudematerial was dried under vacuo, then dissolved in DME/2M Na₂CO₃ (3:1),4-chloro-3-nitropyridine (2.0 equiv.) was added, followed byPd(dppf)Cl₂-DCM (0.1 equiv.). The mixture was heated to 120° C. for 1hr, then EtOAc and H₂O were added, the organic phase was removed, driedwith Na₂SO₄, and concentrated. Purification via SiO₂ columnchromatography eluting with EtOAc yielded6-methyl-3′-nitro-4,4′-bipyridin-2-amine in 35% yield. LCMS (m/z): 231.1(MH⁺) LC R_(t)=1.47 min.

Synthesis of 6-ethyl-N,N-di-BOC-3′-nitro-4,4′-bipyridin-2-amine

To a solution of 6-methyl-3′-nitro-4,4′-bipyridin-2-amine in THF (0.09M)was added BOC₂O (2.2 equiv.), Et₃N (2.5 equiv.), and DMAP (cat.). After5 hrs, the solution was concentrated and filtered through a plug of SiO₂eluting with EtOAc to yield6-ethyl-N,N-di-BOC-3′-nitro-4,4′-bipyridin-2-amine in >95% yield. LCMS(m/z): 431.1 (MH⁺); LC R_(t)=4.29 min.

Synthesis of 6-ethyl-N²,N²-di-BOC-4,4′-bipyridine-2,3′-diamine

To a solution of 6-methyl-N,N-di-BOC-3′-nitro-4,4′-bipyridin-2-amine inEtOH/EtOAc (1:1, 0.2M) was added Pd/C (10% by weight) and the reactionwas stirred under a H₂ balloon for 18 hrs. Filtered through Celite,washed with EtOAc and concentrated the filtrate to afford6-ethyl-N²,N²-di-BOC-4,4′-bipyridine-2,3′-diamine in >95% yield. LCMS(m/z): 401.0 (MH⁺); LC R_(t)=2.81 min.

Synthesis of3-amino-6-bromo-N-(2′-(di-BOC-amino)-6′-methyl-4,4′-bipyridin-3-yl)picolinamide

Method 27 was followed using6-methyl-N²,N²-di-BOC-4,4′-bipyridine-2,3′-diamine (1.0 equiv.),3-amino-6-bromopicolinic acid (1.0 equiv.), EDC (1.0 equiv.), HOAt (1.0equiv.) in NMP (0.48M) to yield3-amino-6-bromo-N-(2′-(di-BOC-amino)-6′-methyl-4,4′-bipyridin-3-yl)picolinamide(35%). LCMS (m/z): 599.1/601.1 (MH⁺); LC R_(t)=3.69 min.

Synthesis of 4-chloro-6-ethylpyridin-2-amine

To a solution of 4,6-dichloropyridin-2-amine (1.0 equiv.) in THF (0.1M)was added Pd(dppf)Cl₂-DCM (0.1M), K₂CO₃ (3.0 equiv.), and Et₂Zn (1.2equiv.). The reaction was heated to 70° C. for 18 hrs. Upon cooling toroom temperature, NH₄Cl(sat.) was added, the mixture was extracted withEtOAc, dried with Na₂SO₄, and concentrated. The crude material waspurified via SiO₂ column chromatography eluting with DCM/MeOH (2%) toyield 4-chloro-6-ethylpyridin-2-amine in 33% yield. LCMS (m/z): 157.1(MH⁺).

Synthesis of 4-chloro-6-ethyl-N,N-di-BOC-pyridin-2-amine

Method 24 was followed using 4-chloro-6-ethylpyridin-2-amine (1.0equiv.), BOC₂O (2.0 equiv.), and DMAP (cat.) in DCM to yield4-chloro-6-ethyl-N,N-di-BOC-pyridin-2-amine (27% yield). LCMS (m/z):357.1 (MH⁺); LC R_(t)=4.11 min.

Synthesis of6-ethyl-N,N-di-BOC-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine

A solution of 4-chloro-6-ethyl-N,N-di-BOC-pyridin-2-amine (1.0 equiv.),bis(pinacolato)diboron (2.0 equiv.), Pd₂(dba)₃ (0.05 equiv.), PCy₃(0.075 equiv.), KOAc (2.0 equiv.) in dioxane (0.19 M) was degassed bybubbling nitrogen through for 10 min in a round-bottomed flask. Theflask was heated to 90° C. for 3 hours, cooled to room temperature,filtered through activated charcoal and Celite and washed with EtOAc.Upon concentration of the filtrate, a thick dark brown crude6-ethyl-N,N-di-BOC-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amineLCMS (m/z): 367.1 (MH⁺ for the corresponding boronic acid).

Synthesis of3-amino-N-(2′-amino-6′-fluoro-4,4′-bipyridin-3-yl)-6-(2,6-difluorophenyl)picolinamide

A solution of3-amino-N-(2′,6′-difluoro-4,4′-bipyridin-3-yl)-6-(2,6-difluorophenyl)picolinamide(1.0 equiv.) in NMP and NH₄OH (2:3, 0.05M) was heated in the microwaveat 120° C. for 8 min. The mixture was directly purified via reversephase HPLC to afford3-amino-N-(2′-amino-6′-fluoro-4,4′-bipyridin-3-yl)-6-(2,6-difluorophenyl)picolinamideas the TFA salt. LCMS (m/z): 437.1 (MH⁺); LC R_(t)=2.79 min.

Synthesis of5-amino-N-(2′-amino-6′-methyl-4,4′-bipyridin-3-yl)-3′-fluoro-2,2′-bipyridine-6-carboxamide

To a solution of degassed dioxane (0.03M) was added3-amino-6-bromo-N-(2′-(di-BOC-amino)-6′-methyl-4,4′-bipyridin-3-yl)picolinamide(1.0 equiv.), bis(pinacolato)diboron (2.0 equiv.), Pd₂(dba)₃ (0.05equiv.), PCy₃ (0.075 equiv.), and KOAc (3.0 equiv.). The solution washeated to 90° C. for 16 hrs until all starting material was consumed.Filtered the reaction and concentrated the filtrate. The crude was driedunder vacuo, then dissolved in DME/2M Na₂CO₃ (3:1, 0.05M), followed byaddition of 2-bromo-3-fluoropyridine (2.0 equiv.) and Pd(dppf)Cl₂-DCM(0.10 equiv.). The reaction was heated to 100° C. in an oil bath untilconsumption of the boronic ester. Cooled to room temperature, added H₂Oand EtOAc, the organic phase was washed with brine, then dried withNa₂SO₄, and concentrated. The crude material was purified via SiO₂column chromatography eluting with EtOAc and hexanes (1:1) and the pureproduct was concentrated and stirred in DCM/TFA (25%) until completionof the deprotection. The reaction was concentrated to dryness andpurified via reverse phase HPLC to afford5-amino-N-(2′-amino-6′-methyl-4,4′-bipyridin-3-yl)-3′-fluoro-2,2′-bipyridine-6-carboxamide.LCMS (m/z): 416.2 (MH⁺); LC R_(t)=1.77 min.

Method 31 Synthesis of3-amino-N-(4-(6-amino-2-(trifluoromethyl)-pyrimidin-4-yl)pyridin-3-yl)-6-(thiazol-4-yl)picolinamide

A solution of tert-butyl6-(3-(3-amino-6-bromopicolinamido)pyridine-4-yl)-2-(trifluoromethyl)pyrimidin-4-ylcarbamate(1.0 equiv.), 4-(tributylstannyl)thiazole (3.0 equiv.), and Pd(PPh₃)₄(0.10 equiv.) in dioxane (0.10M) was microwaved at a 120° C. for 10 min.The reaction was then purified directly via reverse phase HPLC andlyophilized. The product was then stirred in DCM/TFA (25%) untilcompletion of the deprotection, concentrated and purified via reversephase HPLC and lyophilized to give3-amino-N-(4-(6-amino-2-(trifluoromethyl)pyrimidin-4-yl)pyridin-3-yl)-6-(thiazol-4-yl)-picolinamidein 14% yield as the TFA salt. LCMS (m/z): 459.1 (MH⁺); LC R_(t)=2.49min.

Synthesis of(S)-3-amino-N-(4-(3-aminopiperidin-1-yl)pyridin-3-yl)-6-(oxazol-2-yl)picolinamide

Method 31 was followed using (S)-tert-butyl1-(3-(3-amino-6-bromopicolinamido)pyridin-4-yl)piperidin-3-ylcarbamate(1.0 equiv.), 2-(tributyl-stannyl)oxazole (3.0 equiv.), and Pd(PPh3)₄(0.10 equiv.) in dioxane yielding(S)-3-amino-N-(4-(3-aminopiperidin-1-yl)pyridin-3-yl)-6-(oxazol-2-yl)picolinamidein 55% yield as the TFA salt. LCMS (m/z): 380.1 (MH⁺); LC R_(t)=1.55min.

Synthesis of(S)-3-amino-N-(4-(3-aminopiperidin-1-yl)pyridin-3-yl)-6-cyclopropylpicolinamide

Method 31 was followed using (S)-tert-butyl1-(3-(3-amino-6-bromopicolinamido)pyridin-4-yl)piperidin-3-ylcarbamate(1.0 equiv.), 2-cyclopropyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolante(3.0 equiv.), and Pd(dppf)Cl₂-DCM (0.10 equiv.) at 140° C. for 10 minyielding(S)-3-amino-N-(4-(3-aminopiperidin-1-yl)pyridin-3-yl)-6-cyclopropylpicolinamidein 8% yield. LCMS (m/z): 353.1 (MH⁺); LC R_(t)=1.59 min.

Method 32 Synthesis of3-amino-N-(4-((3R,4R)-3-amino-4-hydroxypiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluorophenyl)picolinamide

3-amino-6-(2,6-difluorophenyl)picolinic acid and tert-butyl(3R,4R)-1-(3-aminopyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamatewere coupled following method 11 (example 249), to yield tert-butyl(3R,4R)-1-(3-(3-amino-6-(2,6-difluorophenyl)picolinamido)pyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamateafter HPLC purification. Alternatively, tert-butyl(3R,4R)-1-(3-(3-amino-6-(2,6-difluorophenyl)picolinamido)pyridin-4-yl)-4-(tert-butyldimethylsilyloxy)-piperidin-3-ylcarbamatecould be obtained starting with tert-butyl(3R,4R)-1-(3-(3-amino-6-bromo-picolinamido)pyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-yl-carbamateand 2,6 difluorophenylboronic acid following the Suzuki procedureoutlined in method 14. The TBDMS deprotection of tert-butyl(3R,4R)-1-(3-(3-amino-6-(2,6-difluorophenyl)picolinamido)pyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-yl-carbamatewas performed with 6N HCl, THF, methanol (1:2:1) at room temperature for2 h. After volatile materials were removed, the crude material wasstirred in 30% TFA in dichloromethane for 2 hours. After volatilematerials were removed in vacuo, purification and lyophilization yielded3-amino-N-(4-((3R,4R)-3-amino-4-hydroxy-piperidin-1-yl)pyridin-3-yl)-6-(2,6-difluorophenyl)picolinamide.HPLC. LCMS (m/z): 441.2 (MH⁺); LC R_(t)=2.03 min.

The following compounds were made using method 32:

Ex- ample Structure Name MH+ LC 648

5-Amino-2-(2,6-difluoro- phenyl)-pyrimidine-4- carboxylic acid(3-hydroxy- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 427.22.18 649

5-Amino-2-(2-fluoro-phenyl)- pyrimidine-4-carboxylic acid(3-hydroxy-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)- amide 409.22.26 650

5-Amino-2-phenyl-pyrimidine- 4-carboxylic acid (3-hydroxy-3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 391.2 2.37 651

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid (3- hydroxy-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)- amide493.2 2.46 652

3-Amino-6-(2,6-difluoro-3- isopropoxy-phenyl)-pyridine- 2-carboxylicacid (3-hydroxy- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide484.3 2.94 653

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid(3-hydroxy-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 426.22.66 654

3-Amino-6-(5-chloro-2-fluoro- 4-methyl-phenyl)-pyridine-2- carboxylicacid (3-hydroxy- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide456.1 3.15 655

3-Amino-6-(3-chloro-2-fluoro- phenyl)-pyridine-2-carboxylic acid(3-hydroxy-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 442.12.96 656

3-Amino-6-(2-fluoro-5-phenyl- carbamoyl-phenyl)-pyridine-2- carboxylicacid (3-hydroxy- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide527.2 2.88 657

3-Amino-6-(5-dimethyl- carbamoyl-2-fluoro-phenyl)- pyridine-2-carboxylicacid (3- hydroxy-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)- amide479.2 2.27 658

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid(3-hydroxy-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 426.22.63 659

3-Amino-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (3-hydroxy-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)- amide 408.22.66 660

3-Amino-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (3-hydroxy-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)- amide 408.22.65 661

3-Amino-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (3-hydroxy-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)- amide 408.22.64 662

5-Amino-2-(2-fluoro-phenyl)- pyrimidine-4-carboxylic acid(3-amino-4-hydroxy-3,4,5,6- tetrahydro-2H-[1,4′]bi-pyridinyl-3′-yl)-amide 424.2 1.67 663

5-Amino-2-(2,6-difluoro- phenyl)-pyrimidine-4- carboxylic acid(3-amino-4- hydroxy-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 442.2 1.57 664

3-Amino-6-(2,6-difluoro-3- isopropoxy-phenyl)-5-fluoro-pyridine-2-carboxylic acid (3- amino-4-hydroxy-3,4,5,6-tetra-hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 517.2 2.48 665

3-Amino-6-(2,6-difluoro-3- isopropoxy-phenyl)-pyridine- 2-carboxylicacid (3-amino-4- hydroxy-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)- amide 499.2 2.38 666

3-Amino-6-(2,6-difluoro-3- isopropoxy-phenyl)-5-fluoro-pyridine-2-carboxylic acid (4- amino-3-hydroxy-3,4,5,6-tetra-hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 517.2 2.47 667

3-Amino-6-(2,6-difluoro-3- isopropoxy-phenyl)-pyridine- 2-carboxylicacid (4-amino-3- hydroxy-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)- amide 499.2 2.36 668

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid (4- amino-3-hydroxy-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 508.3 1.96 669

3-Amino-6-thiazol-2-yl- pyridine-2-carboxylic acid (3-amino-4-hydroxy-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide412.1 1.48 670

5-Amino-2-phenyl-pyrimidine- 4-carboxylic acid (3-amino-4-hydroxy-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)- amide 406.21.72 671

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid (3- amino-4-hydroxy-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 508.3 2.00 672

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid(3-amino-5-hydroxy- 3,4,5,6-tetrahydro-2H-[1,4′]bi-pyridinyl-3′-yl)-amide 441.2 1.89 673

5-Amino-2-(2,6-difluoro- phenyl)-pyrimidine-4- carboxylic acid(3-amino-3- methyl-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide440.1 1.76 674

3-Amino-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (3-amino-3-methyl-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide421.2 2.13 675

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid (3- amino-3-methyl-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 506.2 2.08 676

5-Amino-2-(2-fluoro-phenyl)- pyrimidine-4-carboxylic acid(3-amino-3-methyl-3,4,5,6- tetrahydro-2H-[1,4′]bi-pyridinyl-3′-yl)-amide 422.1 1.73 677

3-Amino-6-(2-fluoro-5- isopropylcarbamoyl-phenyl)- pyridine-2-carboxylicacid (3- amino-5-hydroxy-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 508.1 1.89 678

3-Amino-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (5-amino-3-hydroxy-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl- 3′-yl)-amide423.2 1.87 679

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid(3-amino-4-hydroxy- 3,4,5,6-tetrahydro-2H-[1,4′]bi-pyridinyl-3′-yl)-amide 441.2 2.04 680

3-Amino-6-(2,6-difluoro- phenyl)-pyridine-2-carboxylic acid(3-amino-4-hydroxy- 3,4,5,6-tetrahydro-2H-[1,4′]bi-pyridinyl-3′-yl)-amide 441.2 2.04 681

5-Fluoro-6-phenyl-pyridine-2- carboxylic acid (3-amino-4-hydroxy-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)- amide 408.11.99 682

5-Fluoro-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (3-amino-4-hydroxy-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide426.1 2.01 683

6-(2,6-Difluoro-phenyl)-5- fluoro-pyridine-2-carboxylic acid(3-amino-4-hydroxy- 3,4,5,6-tetrahydro-2H-[1,4′]bi-pyridinyl-3′-yl)-amide 444.1 1.99 684

5-Fluoro-6-(2-fluoro-phenyl)- pyridine-2-carboxylic acid (5-amino-3-hydroxy-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide426.1 1.89 685

3-Amino-6-(2-fluoro-5- propoxy-phenyl)-pyridine-2- carboxylic acid(5-amino-3- hydroxy-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)-amide 481.1 2.39

Method 33 Synthesis of (S)-tert-butyl1-(3-(3-amino-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinamido)pyridin-4-yl)piperidin-3-ylcarbamate

A solution of (S)-tert-butyl1-(3-(3-amino-6-bromo-picolinamido)pyridin-4-yl)piperidin-3-ylcarbamate(1.0 eq), bis(pinacolato)diboron (2.0 equiv.), KOAc (3 equiv.),triscyclohexylphosphine (0.075 eq.) in dioxane (0.16 M) was degassed bybubbling argon through for 10 min at which time Pd₂(dba)₃ (0.05 eq.) wasadded. The glass vessel was sealed and heated at 90° C. for 3 hours,cooled to room temperature, filtered, washed with EtOAc and concentratedto give a thick dark brown crude product which was used as is. LCMS(m/z): 457.2 (MH⁺ for the corresponding boronic acid).

Synthesis of (S)-tert-butyl1-(3-(3-amino-6-bromo-5-fluoropicolinamido)pyridin-4-yl)piperidin-3-ylcarbamate

Following Method 11 of Example 305, (+/−)-tert-butyl1-(3-aminopyridin-4-yl)-4-fluoropiperidin-3-ylcarbamate and3-amino-6-bromo-5-fluoro-picolinic acid were reacted yielding(S)-tert-butyl1-(3-(3-amino-6-bromo-5-fluoro-picolinamido)pyridin-4-yl)piperidin-3-ylcarbamate(40%). LCMS (m/z): 509.0 (MH⁺), LC R_(t)=3.04 min.

Synthesis of (S)-tert-butyl1-(3-(3-amino-5-fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinamido)pyridin-4-yl)piperidin-3-ylcarbamate

Starting with (S)-tert-butyl1-(3-(3-amino-6-bromo-5-fluoro-picolinamido)pyridin-4-yl)piperidin-3-ylcarbamate,method 33 was followed yielding (S)-tert-butyl1-(3-(3-amino-5-fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-picolinamido)pyridin-4-yl)piperidin-3-ylcarbamate.LCMS (m/z): 475.2 (MH⁺ for the corresponding boronic acid); LCR_(t)=2.16 min.

Synthesis of tert-butyl(3R,4R)-1-(3-(3-amino-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinamido)pyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate

Starting with tert-butyl(3R,4R)-1-(3-(3-amino-6-bromo-picolinamido)-pyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate,method 33 was followed yielding tert-butyl(3R,4R)-1-(3-(3-amino-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinamido)pyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate.LCMS (m/z): 587.3 (MH⁺ for the corresponding boronic acid).

Method 34 Synthesis of(S)-5-amino-N-(4-(3-aminopiperidin-1-yl)-pyridin-3-yl)-2,2′-bipyridine-6-carboxamide

A solution of (S)-tert-butyl1-(3-(3-amino-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinamido)pyridin-4-yl)piperidin-3-ylcarbamate(1.0 eq), 2-bromo-pyridine (1.0 eq) and Pd(dppf)Cl₂-DCM (0.10 equiv.) in3:1 dimethoxyethane/2M Na₂CO₃ was heated in the microwave at 110° C. for15 minutes. The organic layer was separated, the volatiles were removedin vacuo and the crude material was purified by RP HPLC to yield theN-Boc product after lyophilization. The Boc group was removed bytreating with 25% TFA/CH₂Cl₂ for 2 hours. After removal of volatiles invacuo, purification by RP HPLC and lyophilization(S)-5-amino-N-(4-(3-aminopiperidin-1-yl)-pyridin-3-yl)-2,2′-bipyridine-6-carboxamidewas obtained (12%). The free base and HCl salt can be obtained asdescribed in Method 9 (Example 115). HPLC. LCMS (m/z): 390.2 (MH⁺); LCR_(t)=1.11 min. HCl salt, ¹H NMR (DMSO_(d6)): δ 10.46 (s, 1H), 9.15 (s,1H), 8.61-8.65 (m, 1H), 8.44-8.47 (m, 1H), 8.34 (d, J=9.0 Hz, 2H),7.90-8.05 (m, 3H), 7.41 (d, J=8.7 Hz, 2H), 7.22-7.33 (m, 3H), 2.75-3.60(m, 5H), 1.20-1.95 (m, 4H).

For compounds prepared using method 34 that contained hydroxylfunctionalities, the silyl protecting groups were removed prior to Bocremoval as described in Method 32.

The following compounds were prepared using method 34:

Ex- ample Structure Name MH+ LC 686

3-Amino-6-(4-cyano-2-fluoro- phenyl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 432.22.15 687

3-Amino-6-[2-fluoro-5-(3- methyl-but-1-enyl)-phenyl]-pyridine-2-carboxylic acid (3- amino-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 475.2 2.83 688

3-Amino-6-thiazol-5-yl- pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 396.1 1.52689

3-Amino-6- [2,6-difluoro-3- (isopropyl-methyl-carbamoyl)-phenyl]-pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetra-hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 524.3 2.28 690

5-Amino-3′-fluoro-[2,2′]bi- pyridinyl-6-carboxylic acid (3-hydroxy-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 409.2 1.72691

5′-Amino[2,2′]bipyridinyl-5,6′- dicarboxylic acid 5-amide 6′-[(3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide] 433.2 1.30692

3-Amino-6-isoquinolin-4-yl- pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 440.2 1.53693

5-Amino-1′-oxy-[2,2′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 406.2 1.37694

3-Amino-6-(3-methoxy-pyrazin- 2-yl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 421.21.77 695

5-Amino-6′-benzyloxy-[2,2′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 496.3 2.77696

5-Amino-5′-methoxy-[2,2′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 420.2 1.38697

5-Amino-3′-methoxy-[2,2′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 420.2 1.29698

5-Amino-5′-methanesulfonyl- [2,2′]bipyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 468.21.70 699

5-Amino-5′-methylsulfanyl- [2,2′]bipyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 436.21.54 700

5-Amino-3′,5′-dichloro-[2,2′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 458.1 2.17701

5-Amino-5′-chloro-3′-methyl- [2,2′]bipyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 438.21.92 702

5-Amino-3′-fluoro-[2,2′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 408.2 1.65703

5-Amino-6′-pyrazol-1-yl- [2,2′]bipyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 456.22.21 704

5′-Amino-[2,2′]bipyridinyl-4,6′- dicarboxylic acid 4-amide 6′-[(3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide] 433.2 1.28705

5,5′-Diamino-6′-methoxy- [2,2′]bipyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 435.21.70 706

6′-Acetyl-5-amino-[2,2′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 432.2 2.06707

5-Amino-3′-fluoro-4′-methyl- [2,2′]bipyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 422.21.48 708

5-Amino-5′-fluoro-6′-methyl- [2,2′]bipyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 422.21.75 709

5-Amino-3′-fluoro-6′-methyl- [2,2′]bipyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 422.21.49 710

5-Amino-6′-chloro-[2,2′]bi- pyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetrahydro- 2H-[1,4′]bipyridinyl-3′-yl)- amide 424.22.06 711

5,6′-Diamino-[2,2′]bipyridinyl- 6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 405.2 1.29 712

5-Amino-6′-fluoro-[2,2′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 408.2 1.89713

5-Amino-5′-chloro-[2,2′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 424.2 1.85714

5-Amino-5′-trifluoromethyl- [2,2′]bipyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 458.22.15 715

5-Amino-4′-trifluoromethyl- [2,2′]bipyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 458.22.08 716

5,3′-Diamino-[2,2′]bipyridinyl- 6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 405.2 1.07 717

5-Amino-4′-ethyl-[2,2′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 418.2 1.44718

5-Amino-6′-trifluoromethyl- [2,2′]bipyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 458.22.40 719

5-Amino-3′-chloro-[2,2′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 424.1 1.81720

5-Amino-3′-chloro-5′-trifluoro- methyl-[2,2′]bipyridinyl-6- carboxylicacid (3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide492.2 2.29 721

5-Amino-3′-trifluoromethyl- [2,2′]bipyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 458.21.94 722

3-Amino-6-(6-methoxy-pyrazin- 2-yl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 421.21.93 723

5-Amino-6′-ethoxy-[2,2′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 434.2 2.05724

5-Amino-4′-methyl-[2,2′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 404.2 1.26725

5-Amino-5′-methyl-[2,2′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 404.2 1.25726

5-Amino-6′-methyl-[2,2′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 404.2 1.14727

5-Amino-5′-fluoro-[2,2′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 408.2 1.64728

3-Amino-6-(2-amino-pyrimidin- 4-yl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 406.11.17 729

3-Amino-6-(5-amino-pyrazin-2- yl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 406.11.37 730

5-Amino-[2,2′]bipyridinyl-6- carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 390.2 1.11 731

3-Amino-6-(5-amino-6- methoxy-pyrazin-2-yl)-pyridine- 2-carboxylic acid(3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 436.21.50 732

3-Amino-6-(2,6-difluoro-3- isopropylcarbamoyl-phenyl)-pyridine-2-carboxylic acid (3- amino-4-hydroxy-3,4,5,6-tetra-hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 526.3 1.91 733

3-Amino-6-(2,6-difluoro-3- isopropylcarbamoyl-phenyl)-pyridine-2-carboxylic acid (3- amino-4-fluoro-3,4,5,6-tetra-hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 528.3 2.02 734

5-Amino-3′-fluoro-[2,2′]bi- pyridinyl-6-carboxylic acid (4-amino-3-hydroxy-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide424.2 1.25 735

5-Amino-3′-fluoro-[2,2′]bi- pyridinyl-6-carboxylic acid (3-amino-4-hydroxy-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide424.2 1.37 736

3-Amino-6-(2,6-difluoro-3- isopropylcarbamoyl-phenyl)-5-fluoro-pyridine-2-carboxylic acid (3-amino-3,4,5,6-tetra-hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 528.2 2.13 737

5-Amino-3,3′-difluoro-4′- methyl-[2,2′]bipyridinyl-6- carboxylic acid(3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 440.21.88 738

3-Amino-5-fluoro-6-(6-propoxy- pyrazin-2-yl)-pyridine-2- carboxylic acid(3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 467.22.34 739

3-Amino-5-fluoro-6-(6- isopropoxy-pyrazin-2-yl)- pyridine-2-carboxylicacid (3- amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide467.2 2.31 740

3-Amino-6-(6-ethoxy-pyrazin- 2-yl)-5-fluoro-pyridine-2- carboxylic acid(3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 453.22.11 741

3Amino-5-fluoro-6-(6- isobutoxy-pyrazin-2-yl)- pyridine-2-carboxylicacid (3- amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide481.2 2.53 742

5,5′-Diamino-6′-(3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi-pyridinyl-3′-ylcarbamoyl)-3,3′- difluoro-[2,2′]bipyridinyl-6- carboxylicacid methyl ester 499.2 1.86 743

5-Amino-3′-chloro-3-fluoro- [2,2′]bipyridinyl-6-carboxylic acid(3-amino-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide 442.21.96 744

5-Amino-3,3′-difluoro-[2,2′]bi- pyridinyl-6-carboxylic acid (3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 426.2 1.75745

5-Amino-3,3′-difluoro-6′- methyl-[2,2′]bipyridinyl-6- carboxylic acid(3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 440.21.82 746

5-Amino-3′-fluoro-[2,2′]bi- pyridinyl-6-carboxylic acid (2′-amino-6′-methyl-[4,4′]bi- pyridinyl-3-yl)-amide 416.2 1.77 747

3-Amino-6-(6-propoxy-pyrazin- 2-yl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 449.22.34 748

3-Amino-6-(6-isopropoxy- pyrazin-2-yl)-pyridine-2- carboxylic acid(3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 449.22.29 749

3-Amino-6-(6-ethoxy-pyrazin-2- yl)-pyridine-2-carboxylic acid(3-amino-3,4,5,6-tetrahydro-2H- [1,4′]bipyridinyl-3′-yl)-amide 435.22.12 750

3-Amino-6-(6-benzyloxy- pyrazin-2-yl)-pyridine-2- carboxylic acid(3-amino- 3,4,5,6-tetrahydro-2H-[1,4′]bi- pyridinyl-3′-yl)-amide 497.12.62

Example 7513-Amino-N-(4-((S)-3-aminopiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluoro-3-hydroxyphenyl)-5-fluoropicolinamide

Method 2 of Example 49 was followed using3-amino-N-(4-((S)-3-aminopiperidin-1-yl)pyridin-3-yl)-6-(3-(benzyloxy)-2,6-difluorophenyl)-5-fluoropicolinamidewith 20 wt % Pd/C in methanol (0.1 M solution). The Boc protectedproduct was purified by preparative HPLC. After volatile materials wereremoved, the crude material was stirred in 30% TFA in dichloromethane.After volatile materials were removed in vacuo,3-amino-N-(4-((S)-3-aminopiperidin-1-yl)pyridin-3-yl)-6-(2,6-difluoro-3-hydroxyphenyl)-5-fluoropicolinamidewas obtained by preparative HPLC. LCMS (m/z): 459.2 (MH⁺); LC R_(t)=2.10min.

Synthesis of methyl 3-amino-6-cyclohexylpicolinate

A solution of methyl 3-amino-6-bromopicolinate (1.0 equiv.), cyclohexylzincbromide 0.5 M solution in THF (1.5 equiv.), andtetrakis(triphenylphosphine)-palladium(0) (0.05 equiv.) was stirred at50° C. for 15 minutes. The reaction was filtered and washed with EtOAc.The organic was washed with H₂O (100 mL), NaCl_((sat)) (50 mL), driedover MgSO₄, and the volatiles were removed in vacuo. The product waspurified on silica utilizing the Isco 0-65% gradient of hexane/EtOAc toyield methyl 3-amino-6-cyclohexylpicolinate (98%). LCMS (m/z): 235.2(MH⁺); LC R_(t)=1.89 min.

Synthesis of 3-amino-6-cyclohexylpicolinic acid

To a solution of methyl 3-amino-6-cyclohexylpicolinate (1.0 equiv) inTHF, at a concentration of 0.5 M, was added 1M LiOH (4.0 equiv). Afterstirring for 4 hours at room temperature, 1 N HCl (4.0 equiv.) was addedand the THF was removed in vacuo. The resulting solid was filtered,rinsed with cold H₂O (3×20 mL) yielding 3-amino-6-cyclohexylpicolinicacid (18%). LCMS (m/z): 221.0 (MH⁺); LC R_(t)=4.1 min

To a solution of(3R,5R)-3-(tert-butyldimethylsilyloxy)-5-fluoropiperidine (1 eq) in 30mL of methanol was added 3.8M HCl in isopropanol (4 eq). The reactionmixture was allowed to stand at room temperature for 3 hours at whichpoint it was concentrated under reduced pressure. The resulting residuewas diluted with 120 mL of EtOAc, washed with sat. aq. sodiumbicarbonate, brine, then dried over anhydrous MgSO₄, filtered, andconcentrated in vacuo. The crude residue was purified by flashchromatography (EtOAc:hexanes=2:1) to give (3R,5R)-benzyl3-fluoro-5-hydroxypiperidine-1-carboxylate, (94%). LC/MS (m/z): 254.2(MH⁺).

Synthesis of (3S,5R)-benzyl 3-azido-5-fluoropiperidine-1-carboxylate

To a solution of (3R,5R)-benzyl3-fluoro-5-hydroxypiperidine-1-carboxylate (1 eq) in 14 mL ofdichloromethane was added triethyl amine (3 eq) and methanesulfonylchloride (1.5 eq) at 0° C. The reaction mixture was allowed to stir atroom temperature for 1.5 hours. The crude mixture was diluted with 120mL of diethyl ether, washed with sat. aq. sodium bicarbonate, brine,then dried over anhydrous MgSO₄, filtered, and concentrated in vacuo.The crude residue was dissolved in 16 mL of NMP. Sodium azide (3.0 eq)was added and the resulting suspension was stirred at 80° C. overnight.The reaction mixture was diluted with 200 mL of EtOAc and 100 mL ofhexanes, washed with water, brine, then dried over anhydrous MgSO₄,filtered, and concentrated in vacuo. The crude residue was purified byflash chromatography (EtOAc:hexanes=1:3) to give the titled compound(90%). LC/MS (m/z): 251.1 (MH⁺-28).

Synthesis of (3S,5R)-benzyl3-(tert-butoxycarbonylamino)-5-fluoropiperidine-1-carboxylate

To a solution of (3S,5R)-benzyl 3-azido-5-fluoropiperidine-1-carboxylate(1 eq) in a mixture of 11 mL of pyridine and 1.5 mL of ammoniumhydroxide was added 1M trimethylphosphine (3 eq) at room temperature.The reaction mixture was stirred at room temperature for 3 hours atwhich point the solvents were removed under reduced pressure to give ayellow oil. The oil was again dissolved in 100 mL of ethanol andconcentrated to remove ammonium hydroxide completely. The residue wasdissolved in 12 ml of 1,4-dioxane and 12 mL of sat. aq. NaHCO₃ wasadded. Di-tert-butyl dicarbonate (4 eq) in 6 mL of THF was addeddropwise at 0° C. The mixture was allowed to stir at room temperaturefor 1 hour. The crude mixture was diluted with 150 mL of EtOAc, washedwith brine, then dried over anhydrous MgSO₄, filtered, and concentratedin vacuo. The crude residue was purified by flash chromatography(EtOAc:hexanes=1:1) to give the titled compound (95%). LC/MS (m/z):253.1 (MH⁺-100).

Synthesis of tert-butyl(3S,5R)-5-fluoro-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate

To a solution of (3S,5R)-benzyl3-(tert-butoxycarbonylamino)-5-fluoropiperidine-1-carboxylate (1 eq) in28 methanol was added 10% Pd/C (0.1 eq). The resulting suspension wasstirred at H₂ atmosphere for 1 hours. The crude solids were filteredthrough a pad of Celite on a paper lined Buchner funnel, washed withMeOH, then concentrated in vacuo. The residue was dissolved in 33 mL ofisopropanol and DIPEA (2.5 eq) and 4-chloro-3-nitropyridine (1.5 eq)were added. The reaction mixture was stirred at 80° C. for 2 hours, atwhich point the reaction mixture was allowed to cool to room temperatureand concentrated under reduced pressure. The residue was diluted with150 mL of EtOAc, washed with brine, then dried over anhydrous MgSO₄,filtered, and concentrated in vacuo. The crude residue was purified byflash chromatography (5% methanol in EtOAc:hexanes=1:1) to give thetitled compound (90%). LC/MS (m/z): 341.1 (MH⁺). HPLC: R_(t): 2.115 min.

Synthesis of tert-Butyl(3S,5R)-1-(3-aminopyridin-4-yl)-5-fluoropiperidin-3-ylcarbamate

Following Method 2 of Example 49, tert-butyl(3S,5R)-5-fluoro-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate wasreduced yielding tert-Butyl(3S,5R)-1-(3-aminopyridin-4-yl)-5-fluoropiperidin-3-ylcarbamate. LC/MS(m/z): 311.1 (MH⁺).

Synthesis of tert-butyl(3S,5R)-1-(3-(3-amino-6-bromopicolinamido)pyridin-4-yl)-5-fluoropiperidin-3-ylcarbamate

Following Method 11 of Example 305, tert-Butyl(3S,5R)-1-(3-aminopyridin-4-yl)-5-fluoropiperidin-3-ylcarbamate and3-amino-6-bromopicolinic acid were coupled yielding after columnchromatography tert-butyl(3S,5R)-1-(3-(3-amino-6-bromopicolinamido)pyridin-4-yl)-5-fluoropiperidin-3-ylcarbamate.LC/MS (m/z): 509.1/511.1 (MH⁺).

Synthesis of cis (+/−)-1-benzyl 3-methyl5-(tert-butoxycarbonylamino)piperidine-1,3-dicarboxylate

To a solution of cis(+/−)-1-(benzyloxycarbonyl)-5-(tert-butoxycarbonylamino)piperidine-3-carboxylicacid (1.0 eq), methanol (20 eq.) and EDC (1.3 eq) in dichloromethane ata concentration of 0.25 M at 0° C. was added dimethylaminopyridine (0.1eq). After stirring for 48 hours as the reaction was allowed to warm tort the volatiles were removed in vacuo. Upon addition of ethyl actetateand washing with H₂O (3×), 1N HCl, NaHCO_(3 (sat.)) and brine, thesolution was dried over MgSO₄, filtered, concentrated and purified bycolumn chromatography (25% ethyl acetate/hexanes) to yield cis(+/−)-1-benzyl 3-methyl5-(tert-butoxycarbonylamino)piperidine-1,3-dicarboxylate. LCMS (m/z):293.1 (MH-Boc⁺); LC R_(t)=4.09 min

Synthesis of cis (+/−)-benzyl3-(tert-butoxycarbonylamino)-5-(hydroxymethyl)piperidine-1-carboxylate

A solution of cis (+/−)-1-benzyl 3-methyl5-(tert-butoxycarbonylamino)piperidine-1,3-dicarboxylate (1.0 eq.) inTHF at a concentration of 0.08 M was cooled at 0° C. and then LiCl (2.3eq.) and sodium borohydride (2.3 eq.) were added. After stirring for 20hours as the reaction warmed to rt, the pH was adjusted with 1M citricacid to pH 4-5. After removal of the volatiles in vacuo, the product wasextracted in dichloromethane, washed with H₂O and brine, dried overMgSO₄. Upon filtering and removal of the volatiles in vacuo, cis(+/−)-benzyl3-(tert-butoxycarbonylamino)-5-(hydroxymethyl)piperidine-1-carboxylatewas obtained as a white foamy solid. LCMS (m/z): 265.0 (MH-Boc⁺); LCR_(t)=3.37 min.

Synthesis of cis (+/−)-benzyl3-(tert-butoxycarbonylamino)-5-((tert-butyldimethylsilyloxy)methyl)piperidine-1-carboxylate

A solution of cis (+/−)-benzyl3-(tert-butoxycarbonylamino)-5-(hydroxymethyl)piperidine-1-carboxylate(1.0 eq.), imidazole (1.1 eq.), tert-butyl-dimethylsilylchloride (1.1eq.) and dimethylaminopyridine (0.1 eq.) in dichloromethane at aconcentration of 0.1 M was stirred for 18 hours at which time thevolatiles were removed in vacuo. Direct purification of the crudematerial by column chromatography (20% ethyl acetate/hexanes) yieldedcis (+/−)-benzyl3-(tert-butoxycarbonylamino)-5-((tert-butyldimethylsilyloxy)methyl)piperidine-1-carboxylate.LCMS (m/z): 379.0 (MH-Boc⁺); LC R_(t)=5.95 min.

Synthesis of cis (+/−)-tert-butyl5-((tert-butyldimethylsilyloxy)-methyl)piperidin-3-ylcarbamate

Method 17 was followed to deprotect cis (+/−)-benzyl3-(tert-butoxycarbonylamino)-5-((tert-butyldimethylsilyloxy)methyl)piperidine-1-carboxylateyielding cis (+/−)-tert-butyl5-((tert-butyldimethylsilyloxy)methyl)piperidin-3-ylcarbamate. LCMS(m/z): 344.1 (MH⁺).

Synthesis of cis (+/−)-tert-butyl5-((tert-butyldimethylsilyloxy)methyl)-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate

Method 1 of Example 1 was followed using cis (+/−)-tert-butyl5-((tert-butyldimethylsilyloxy)methyl)piperidin-3-ylcarbamate and4-chloro-3-nitropydidine yielding cis (+/−)-tert-butyl5-((tert-butyldimethylsilyloxy)methyl)-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate.LCMS (m/z): 467.0 (MH⁺); LC R_(t)=4.02 min.

Synthesis of cis (+/−)-tert-butyl1-(3-aminopyridin-4-yl)-5-((tert-butyldimethylsilyloxy)methyl)piperidin-3-ylcarbamate

Following Method 2 of Example 49, cis (+/−)-tert-butyl5-((tert-butyldimethylsilyloxy)methyl)-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamatewas reduced yielding cis (+/−)-tert-butyl1-(3-aminopyridin-4-yl)-5-((tert-butyldimethylsilyloxy)methyl)piperidin-3-ylcarbamate.LCMS (m/z): 437.2 (MH⁺); LC R_(t)=3.86 min.

Synthesis of cis (+/−)-tert-butyl1-(3-(3-amino-6-bromopicolinamido)pyridin-4-yl)-5-(hydroxymethyl)piperidin-3-ylcarbamate

Following Method 11 of Example 305, cis (+/−)-tert-butyl1-(3-aminopyridin-4-yl)-5-((tert-butyldimethylsilyloxy)methyl)piperidin-3-ylcarbamateand 6-bromo-3-aminopicolinic acid were coupled. Following purificationby RP HPLC the product fractions were allowed to stand at rt overnightin the 0.1% TFA acetonitrile/water solution which removed the silylgroup. Upon subsequent lyophilization, cis (+/−)-tert-butyl1-(3-(3-amino-6-bromopicolinamido)pyridin-4-yl)-5-(hydroxymethyl)piperidin-3-ylcarbamatewas obtained and used in Suzuki reactions directly. LCMS (m/z):521.0/523.1 (MH⁺); LC R_(t)=2.58 min.

Synthesis of cis (+/−)-tert-butyl1-(3-(6-bromo-3-fluoropicolinamido)pyridin-4-yl)-5-(hydroxymethyl)piperidin-3-ylcarbamate

Following Method 11 of Example 305, cis (+/−)-tert-butyl1-(3-aminopyridin-4-yl)-5-((tert-butyldimethylsilyloxy)methyl)piperidin-3-ylcarbamateand 6-bromo-3-fluoropicolinic acid were coupled. Following purificationby RP HPLC the product fractions were allowed to stand at rt overnightin the 0.1% TFA acetonitile/water solution which removed the silylgroup. Upon subsequent lyophilization, cis (+/−)-tert-butyl1-(3-(6-bromo-3-fluoropicolinamido)pyridin-4-yl)-5-(hydroxymethyl)piperidin-3-ylcarbamatewas obtained and used in Suzuki reactions directly. LCMS (m/z):524.0/526.0 (MH⁺); LC R_(t)=2.90 min.

Synthesis of cis (+/−)-benzyl3-(tert-butoxycarbonylamino)-5-(fluoromethyl)piperidine-1-carboxylate

A solution of cis (+/−)-benzyl3-(tert-butoxycarbonylamino)-5-(hydroxymethyl)piperidine-1-carboxylate(1 eq.), perfluorobutanesulfonylfluoride (2 eq.), triethylamine-HF (4eq.) and triethylamine (6 eq.) in tetrahydrofuran at a concentration of0.16 M was stirred for 36 hours. Upon dilution with ethyl acetate (50×)the solution was washed with 1N HCl, NaHCO_(3 (sat)) and brine, wasdried over MgSO₄, filtered, concentrated and purified by columnchromatography (25-40% ethyl acetate/hexanes) to yield cis (+/−)-benzyl3-(tert-butoxycarbonylamino)-5-(fluoromethyl)piperidine-1-carboxylate(45% yield). LCMS (m/z): 267.1 (MH⁺); LC R_(t)=4.23 min.

Synthesis of cis (+/−)-tert-butyl5-(fluoromethyl)piperidin-3-ylcarbamate

Method 17 was followed to deprotect cis (+/−)-benzyl3-(tert-butoxycarbonylamino)-5-(fluoromethyl)piperidine-1-carboxylateyielding cis (+/−)-tert-butyl 5-(fluoromethyl)piperidin-3-ylcarbamate.LCMS (m/z): 233.1 (MH⁺).

Synthesis of cis (+/−)-tert-butyl5-(fluoromethyl)-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate

Method 1 of Example 1 was followed using cis (+/−)-tert-butyl5-(fluoromethyl)piperidin-3-ylcarbamate and 4-chloro-3-nitropyridineyielding cis (+/−)-tert-butyl5-(fluoromethyl)-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate. LCMS(m/z): 355.1 (MH⁺); LC R_(t)=2.41 min.

Synthesis of cis (+/−)-tert-butyl1-(3-aminopyridin-4-yl)-5-(fluoromethyl)piperidin-3-ylcarbamate

Following Method 2 of Example 49, cis (+/−)-tert-butyl5-(fluoromethyl)-1-(3-nitropyridin-4-yl)piperidin-3-ylcarbamate wasreduced yielding cis (+/−)-tert-butyl1-(3-aminopyridin-4-yl)-5-(fluoromethyl)piperidin-3-ylcarbamate. LCMS(m/z): 325.1 (MH⁺); LC R_(t)=2.27 min.

Synthesis of tert-butyl(3R,4R)-1-(3-(6-bromo-5-fluoropicolinamido)pyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate

Following Method 11 of Example 305, tert-butyl(3R,4R)-1-(3-aminopyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamateand 6-bromo-5-fluoropicolinic acid were coupled to yield tert-butyl(3R,4R)-1-(3-(6-bromo-5-fluoropicolinamido)pyridin-4-yl)-4-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate.LCMS (m/z): 510.0/512.0 (MH⁺); LC R_(t)=4.51 min.

Synthesis of tert-butyl(3S,5R)-1-(3-(6-bromo-5-fluoropicolinamido)pyridin-4-yl)-5-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate

Following Method 11 of Example 305, tert-butyl(3S,5R)-1-(3-aminopyridin-4-yl)-5-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamateand 6-bromo-5-fluoropicolinic acid were coupled to yield tert-butyl(3S,5R)-1-(3-(6-bromo-5-fluoropicolinamido)pyridin-4-yl)-5-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate.LCMS (m/z): 624.1/626.1 (MH⁺).

Synthesis of tert-butyl(3S,5R)-1-(3-(3-amino-6-bromopicolinamido)pyridin-4-yl)-5-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate

Following Method 11 of Example 305, tert-butyl(3S,5R)-1-(3-aminopyridin-4-yl)-5-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamateand 6-bromo-3-aminopicolinic acid were coupled to yield tert-butyl(3S,5R)-1-(3-(3-amino-6-bromopicolinamido)pyridin-4-yl)-5-(tert-butyldimethylsilyloxy)piperidin-3-ylcarbamate.LCMS (m/z): 621.1/623.2 (MH⁺).

Synthesis of (3R,4R)-benzyl3-(tert-butoxycarbonylamino)-4-(methylsulfonyloxy)piperidine-1-carboxylate

To a solution of (3R,4R)-benzyl3-(tert-butoxycarbonylamino)-4-hydroxypiperidine-1-carboxylate indichloromethane (0.13 M) was added triethylamine (1.5 equiv.) followedby methanesulfonyl chloride (1.3 equiv.). The reaction was allowed tostir at room temperature for 15 h. The solution was then quenched withsaturated NaHCO₃, extracted with dichloromethane, dried with sodiumsulfate, and concentrated to give the crude product in >95% yield. LCMS(m/z): 428.9/328.9 (MH⁺), LC R_(t)=3.81 min.

Synthesis of (3aR,7aS)-benzyl2-oxohexahydrooxazolo[4,5-c]pyridine-5(6H)-carboxylate

A solution of (3R,4R)-benzyl3-(tert-butoxycarbonylamino)-4-(methylsulfonyloxy)piperidine-1-carboxylatein pyridine (0.16 M) was heated to 120° C. in the microwave for 10minutes. The solution was then concentrated to almost dryness and theforming solid was filtered to give the desired product. The filtrate wasfurther purified via silica gel column chromatography eluting with ethylacetate (100%) to give the product in 75% combined yield. LCMS (m/z):277.1 (MH⁺), LC R_(t)=2.33 min.

Synthesis of (3S,4S)-benzyl3-(tert-butoxycarbonylamino)-4-(methylsulfonyloxy)piperidine-1-carboxylate

To a solution of (3S,4S)-benzyl3-(tert-butoxycarbonylamino)-4-hydroxypiperidine-1-carboxylate indichloromethane (0.13 M) was added triethylamine (1.5 equiv.) followedby methanesulfonyl chloride (1.3 equiv.). The reaction was allowed tostir at room temperature for 15 h. The solution was then quenched withsaturated NaHCO₃, extracted with dichloromethane, dried with sodiumsulfate, and concentrated to give the crude product in >95% yield. LCMS(m/z): 428.9/328.9 (MH⁺), LC R_(t)=3.81 min.

Synthesis of (3aS,7aR)-benzyl2-oxohexahydrooxazolo[4,5-c]pyridine-5(6H)-carboxylate

The previously described method for the enantiomeric compound wasfollowed using (3S,4S)-benzyl3-(tert-butoxycarbonylamino)-4-(methylsulfonyloxy)-piperidine-1-carboxylate(1.0 equiv.) to yield (3aS,7aR)-benzyl2-oxohexahydro-oxazolo[4,5-c]pyridine-5(6H)-carboxylate in 62% yield.LCMS (m/z): 277.1 (MH⁺), LC R_(t)=2.33 min.

Synthesis of (3aR,7aS)-5-benzyl 3-tert-butyl2-oxotetrahydrooxazolo[4,5-c]pyridine-3,5(2H,6H)-dicarboxylate

To a solution of (3aR,7aS)-benzyl2-oxohexahydrooxazolo[4,5-c]pyridine-5(6H)-carboxylate (1.0 equiv.) indichloromethane (0.09 M) was added BOC₂O (1.1 equiv.), triethylamine(1.1 equiv.), and a catalytic amount of DMAP. The reaction was stirredat room temperature for one hour at which point it was concentratedunder vacuo and filtered through a plug of silica gel eluting withethylacetate. The product was dried under vacuo to give a white solid in75% yield. LCMS (m/z): 277.2 (MH⁺), LC R_(t)=3.43 min.

Synthesis of (3aS,7aR)-5-benzyl 3-tert-butyl2-oxotetrahydrooxazolo[4,5-c]pyridine-3,5(2H,6H)-dicarboxylate

The previously described method for the enantiomeric compound wasfollowed using (3aS,7aR)-benzyl2-oxohexahydrooxazolo[4,5-c]pyridine-5(6H)-carboxylate (1.0 equiv.) toyield (3aS,7aR)-5-benzyl 3-tert-butyl2-oxotetra-hydrooxazolo[4,5-c]pyridine-3,5(2H,6H)-dicarboxylate in 90%yield. LCMS (m/z): 277.2 (MH⁺), LC R_(t)=3.43 min.

Synthesis of (3aR,7aS)-tert-butyl5-(3-nitropyridin-4-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-3(2H)-carboxylate

Following Method 17, the Cbz group of (3aR,7aS)-5-benzyl 3-tert-butyl2-oxotetrahydrooxazolo[4,5-c]pyridine-3,5(2H,6H)-dicarboxylate wasremoved and the resulting amine was reacted with4-chloro-3-nitropyridine following Method 1 to yield (3aR,7aS)-tert-butyl 5-(3-nitropyridin-4-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-3(2H)-carboxylate as a yellow foam in 89% yield.LCMS (m/z): 365.1 (MH⁺), LC R_(t)=1.79 min.

Synthesis of (3aS,7aR)-tert-butyl5-(3-nitropyridin-4-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-3(2H)-carboxylate

The previously described method for the enantiomeric compound wasfollowed using (3aS,7aR)-5-benzyl 3-tert-butyl2-oxotetrahydrooxazolo[4,5-c]pyridine-3,5(2H,6H)-dicarboxylate (1.0equiv.) to yield (3aS,7aR)-tert-butyl5-(3-nitropyridin-4-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-3(2H)-carboxylatein 88% yield. LCMS (m/z): 365.1 (MH⁺), LC R_(t)=1.79 min.

Synthesis of (3aR,7aS)-tert-butyl5-(3-aminopyridin-4-yl)-2-oxohexahydro-oxazolo[4,5-c]pyridine-3(2H)-carboxylate

Following Method 2 of Example 49, (3aR,7aS)-tert-butyl5-(3-nitropyridin-4-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-3(2H)-carboxylatein EtOH and EtOAc (1:1, 0.15 M) was reduced yielding(3aR,7aS)-tert-butyl5-(3-aminopyridin-4-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-3(2H)-carboxylatein >95% yield. LCMS (m/z): 335.0 (MH⁺), LC R_(t)=1.68 min.

Synthesis of (3aS,7aR)-tert-butyl5-(3-aminopyridin-4-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-3(2H)-carboxylate

The previously described method for the enantiomeric compound wasfollowed yielding (3aS,7aR)-tert-butyl5-(3-aminopyridin-4-yl)-2-oxohexahydro-oxazolo[4,5-c]pyridine-3(2H)-carboxylatein 97% yield. LCMS (m/z): 335.0 (MH⁺), LC R_(t)=1.68 min.

Synthesis of (3aR,7aS)-tert-butyl5-(3-(3-amino-6-(2,6-difluorophenyl)picolinamido)-pyridin-4-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-3(2H)-carboxylate

To a solution of (3aR,7aS)-tert-butyl5-(3-aminopyridin-4-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-3(2H)-carboxylate(1.0 equiv.) in DMF (0.3 M) was added3-amino-6-(2,6-difluorophenyl)picolinic acid (1.2 equiv.), EDC (1.2equiv.) and HOAt (1.2 equiv.). The solution was stirred for 15 h. To themixture was added water and the precipitate was filtered. To thefiltrate was added EtOAc, and the organic solution was extracted (3times), dried with Na₂SO₄, and concentrated to give an orange syrup. Thecrude was triturated with EtOAc and hexanes mixture and the precipitatewas filtered off to give pure product in 46% yield. LCMS (m/z): 567.0(MH⁺), LC R_(t)=3.03 min.

Synthesis of (3aS,7aR)-tert-butyl5-(3-(3-amino-6-(2,6-difluorophenyl)picolinamido)pyridin-4-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-3(2H)-carboxylate

The previously described method for the enantiomeric compound wasfollowed using (3aS,7 aR)-tert-butyl5-(3-aminopyridin-4-yl)-2-oxohexahydro-oxazolo[4,5-c]pyridine-3(2H)-carboxylateto give (3aS,7 aR)-tert-butyl 5-(3-(3-amino-6-(2,6-di fluorophenyl)p icolinamido)pyridin-4-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-3(2H)-carboxylate. LCMS (m/z): 567.0 (MH⁺),R_(t)=2.86 min.

Synthesis of (3aR,7aS)-tert-butyl5-(3-(3-amino-6-(2-fluoro-5-(isopropylcarbamoyl)phenyl)picolinamido)pyridin-4-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-3(2H)-carboxylate

To a solution of (3aR,7aS)-tert-butyl5-(3-aminopyridin-4-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-3(2H)-carboxylate(1.0 equiv.) in DMF (0.3 M) was added3-amino-6-(2-fluoro-5-(isopropylcarbamoyl)phenyl)picolinic acid (1.2equiv.), EDC (1.2 equiv.) and HOAt (1.2 equiv.). The solution wasstirred for 15 h. To the mixture was added water and the precipitate wasfiltered. To the filtrate was added EtOAc, and the organic solution wasextracted (3 times), dried with Na₂SO₄, and concentrated to give anorange syrup. The crude was triturated with EtOAc and hexanes mixtureand the precipitate was filtered off to give (3aR,7aS)-tert-butyl5-(3-(3-amino-6-(2-fluoro-5-(isopropylcarbamoyl)phenyl)picolinamido)pyridin-4-yl)-2-oxohexahydro-oxazolo[4,5-c]pyridine-3(2H)-carboxylate.LCMS (m/z): 634.3.

Method 35 Synthesis of3-amino-N-(4-((3R,4S)-3-amino-4-hydroxypiperidin-1-yl)-pyridin-3-yl)-6-(2,6-difluorophenyl)picolinamide

To a solution of (3aR,7aS)-tert-butyl5-(3-(3-amino-6-(2,6-difluoro-phenyl)picolinamido)pyridin-4-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-3(2H)-carboxylatein MeOH (0.06M) was added Cs₂CO₃ (0.5 equiv.) and the reaction wasstirred for 3 h. The mixture was then concentrated to dryness undervacuo and the crude was stirred in TFA and DCM (25% TFA) untilcompletion. The reaction was concentrated and purified via reverse phaseHPLC. Upon lyophilization, a white powder was obtained as the TFA salt.LCMS (m/z): 441.1 (MH⁺), LC R_(t)=1.95 min.

The following compounds were prepared using Method 35:

Ex- ample Structure Name MH+ LC 752

3-Amino-6-(2,6-difluoro-phenyl)- pyridine-2-carboxylic acid (3-amino-4-hydroxy-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide441.0 1.98 753

3-Amino-6-(2-fluoro-5-isopropyl- carbamoyl-phenyl)-pyridine-2-carboxylic acid (3-amino-4- hydroxy-3,4,5,6-tetrahydro-2H-[1,4′]bipyridinyl-3′-yl)-amide 508.0 1.98 754

3-Amino-6-(2,6-difluoro-phenyl)- pyridine-2-carboxylic acid (3-amino-4-hydroxy-3,4,5,6-tetra- hydro-2H-[1,4′]bipyridinyl-3′- yl)-amide441.1 1.95

Example 755 Synthesis of(S)-3-amino-N-(4-(3-aminopiperidin-1-yl)pyridin-3-yl)-6-(2-fluoro-5-isopentylphenyl)picolinamide

Method 2 of Example 49 was followed using (S,Z)-tert-butyl1-(3-(3-amino-6-(2-fluoro-5-(3-methylbut-1-enyl)phenyl)picolinamido)pyridin-4-yl)piperidin-3-ylcarbamatewhich after Boc deprotection with 25% TFA/CH₂Cl₂ yielded(S)-3-amino-N-(4-(3-aminopiperidin-1-yl)pyridin-3-yl)-6-(2-fluoro-5-isopentylphenyl)picolinamide(35%). LCMS (m/z): 477.3 (MH⁺); LC R_(t)=2.91 min.

Example 756 Pim1 ATP Depletion Assay

The activity of PIM1 is measured using a luciferase-luciferin based ATPdetection reagent to quantify ATP depletion resulting fromkinase-catalyzed phosphoryl transfer to a peptide substrate. Compoundsto be tested are dissolved in 100% DMSO and directly distributed intowhite 384-well plates at 0.5 μl per well. To start the reaction, 10 μlof 5 nM Pim1 kinase and 80 μM BAD peptide (RSRHSSYPAGT-OH) in assaybuffer (50 mM HEPES pH 7.5, 5 mM MgCl₂, 1 mM DTT, 0.05% BSA) is addedinto each well. After 15 minutes, 10 μl of 40 μM ATP in assay buffer isadded. Final assay concentrations are 2.5 nM PIM1, 20 μM ATP, 40 μM BADpeptide and 2.5% DMSO. The reaction is performed until approximately 50%of the ATP is depleted, then stopped with the addition of 20 μlKinaseGlo Plus (Promega Corporation) solution. The stopped reaction isincubated for 10 minutes and the remaining ATP detected via luminescenceon the Victor2 (Perkin Elmer). Compounds of the foregoing examples weretested by the Pim1 ATP depletion assay and found to exhibit an IC₅₀values as shown in Example 763, below. IC₅₀, the half maximal inhibitoryconcentration, represents the concentration of a test compound that isrequired for 50% inhibition of its target in vitro.

Example 757 Pim2 ATP Depletion Assay

The activity of PIM2 is measured using a luciferase-luciferin based ATPdetection reagent to quantify ATP depletion resulting fromkinase-catalyzed phosphoryl transfer to a peptide substrate. Compoundsto be tested are dissolved in 100% DMSO and directly distributed intowhite 384-well plates at 0.5 μl per well. To start the reaction, 10 μlof 10 nM Pim2 kinase and 20 μM BAD peptide (RSRHSSYPAGT-OH) in assaybuffer (50 mM HEPES pH 7.5, 5 mM MgCl₂, 1 mM DTT, 0.05% BSA) is addedinto each well. After 15 minutes, 10 μl of 8 μM ATP in assay buffer isadded. Final assay concentrations are 5 nM PIM2, 4 μM ATP, 10 μM BADpeptide and 2.5% DMSO. The reaction is performed until approximately 50%of the ATP is depleted, then stopped with the addition of 20 μlKinaseGlo Plus (Promega Corporation) solution. The stopped reaction isincubated for 10 minutes and the remaining ATP detected via luminescenceon the Victor2 (Perkin Elmer). Compounds of the foregoing examples weretested by the Pim2 ATP depletion assay and found to exhibit an IC₅₀values as shown in Example 763, below.

Example 758 Pim3 ATP Depletion Assay

The activity of PIM3 is measured using a luciferase-luciferin based ATPdetection reagent to quantify ATP depletion resulting fromkinase-catalyzed phosphoryl transfer to a peptide substrate. Compoundsto be tested are dissolved in 100% DMSO and directly distributed intowhite 384-well plates at 0.5 μl per well. To start the reaction, 10 μlof 10 nM Pim3 kinase and 200 μM BAD peptide (RSRHSSYPAGT-OH) in assaybuffer (50 mM HEPES pH 7.5, 5 mM MgCl₂, 1 mM DTT, 0.05% BSA) is addedinto each well. After 15 minutes, 10 μl al of 80 μM ATP in assay bufferis added. Final assay concentrations are 5 nM PIM1, 40 μM ATP, 100 μMBAD peptide and 2.5% DMSO. The reaction is performed until approximately50% of the ATP is depleted, then stopped by the addition of 20 μlKinaseGlo Plus (Promega Corporation) solution. The stopped reaction isincubated for 10 minutes and the remaining ATP detected via luminescenceon the Victor2 (Perkin Elmer). Compounds of the foregoing examples weretested by the Pim3 ATP depletion assay and found to exhibit an IC₅₀values as shown in Example 763, below.

Example 759 Flt3 AlphaScreen Assay

The activity of Flt3 is measured using a homogeneous bead based systemquantifying the amount of phosphorylated peptide substrate resultingfrom kinase-catalyzed phosphoryl transfer to a peptide substrate.Compounds to be tested are dissolved in 100% DMSO and directlydistributed into white 384-well plates at 0.5 μl per well. To start thereaction, 10 μl of 300 μM Flt3 kinase and 700 μM ATP in assay buffer (50mM Hepes, pH=7.5, 5 mM MgCl₂, 0.05% BSA, 1 mM DTT) is added into eachwell followed by 10 μl of 500 nM SHC peptide(Biotin-GGLFDDPSYVNVQNL-NH₂) in assay buffer. Final assay concentrationsare 150 μM Flt3, 350 μM ATP, 250 nM SHC peptide and 2.5% DMSO. Thereaction is performed for 2.5 hours, then stopped by the addition of 10μl 60 mM EDTA. 25 μl of 1.2 μg/ml PY20 antibody, 48.4 μg/ml Protein AAlpha Screen beads, and 48.4 μg/ml streptavidin coated Alpha Screenbeads in detection buffer (50 mM Tris pH 7.5, 0.01% Tween-20) is addedto the stopped reactions. The stopped reactions are incubated overnightin the dark. The phosphorylated peptide is detected via an oxygen anioninitiated chemiluminescence/fluorescence cascade using the Envisionplate reader (Perkin Elmer). Compounds of the foregoing examples weretested by the Flt3 Alpha assay and found to exhibit an IC₅₀ values asshown in Example 762, below.

Example 760 KDR AlphaScreen Assay

The activity of KDR is measured using a homogeneous bead based systemquantifying the amount of phosphorylated peptide substrate resultingfrom kinase-catalyzed phosphoryl transfer to a peptide substrate.Compounds to be tested are dissolved in 100% DMSO and directlydistributed to a white 384-well plate at 0.5 μl per well. To start thereaction, 10 μl of 2 μM VEGFS peptide (Biotin-GGGGQDGKDYIVLP1—NH₂) inassay buffer (50 mM Hepes, pH=7.5, 5 mM MnCl₂, 0.1% BSA, 0.01% Tween-20,1 mM DTT) is added to each well followed by the addition of 10 μl of 250μM KDR kinase and 2 μM ATP in assay buffer. Final assay concentrationsare 125 μM KDR, 1 μM ATP, 1 μM VEGFS peptide and 2.5% DMSO. The reactionis performed for 2 hours, then stopped by the addition of 25 μl of 0.24μg/ml PY20 antibody, 96.8 μg/ml Protein A Alpha Screen beads, and 96.8μg/ml streptavidin coated Alpha Screen beads in stop/detection buffer(50 mM Hepes, pH=7.5, 10 mM EDTA, 0.1% BSA, 0.01% Tween-20). The stoppedreactions are incubated overnight in the dark. The phosphorylatedpeptide is detected via an oxygen anion initiatedchemiluminescence/fluorescence cascade using the Envision plate reader(Perkin Elmer). Compounds of the foregoing examples were tested by theKDR Alpha Screen assay and found to exhibit an IC₅₀ values as shown inExample 762, below.

Example 761 Cell Proliferation Assay

HEL 92.1.7 cells (ATTC No. TIB-180, an erythroleukemia line derived frommalignant peripheral blood), MV4-11 cells (ATCC No. CRL-9591, a humanacute monocytic leukemia line) and PC3 cells (ATCC No. CRL-1435, a humanprostatic adenocarcinoma line) were cultured in RPMI1640 supplementedwith 10% FBS, sodium pyruvate and antibiotics. Cells were plated in thesame medium at a density of 1000 cells per well into 96 well tissueculture plates, with outside wells vacant, on the day of assay.

KMS 11 (human myeloma cell line), were cultured in IMDM supplementedwith 10% FBS, sodium pyruvate and antibiotics. Cells were plated in thesame medium at a density of 2000 cells per well into 96 well tissueculture plates, with outside wells vacant, on the day of assay. MM1.s(human myeloma cell line), were cultured in RPMI1640 supplemented with10% FBS, sodium pyruvate and antibiotics. Cells were plated in the samemedium at a density of 5000 cells per well into 96 well tissue cultureplates, with outside wells vacant, on the day of assay.

Test compounds supplied in DMSO were diluted into DMSO at 500 times thedesired final concentrations before dilution into culture media to 2times final concentrations. Equal volumes of 2× compounds were added tothe cells in 96 well plates and incubated at 37° C. for 3 days.

After 3 days plates were equilibrated to room temperature and equalvolume of CellTiter-Glow Reagent (Promega) was added to the culturewells. The plates were agitated briefly and luminescent signal wasmeasured with luminometer. The percent inhibition of the signal seen incells treated with DMSO alone vs. cells treated with control compoundwas calculated and used to determine EC₅₀ values (i.e., theconcentration of a test compound that is required to obtain 50% of themaximum effect in the cells) for tested compounds, as shown in Example763.

Example 762 In Vitro PKCE Assay

An in vitro PKCE assay was run using 10 nM final concentration of humanfull length PKCE enzyme purchased from InVitrogen. Peptide substratewith the sequence ERMRPRKRQGSVRRRV-OH was used at final concentration of40 uM, and ATP at 20 uM. Lipid activator, 0.05 mg/ml ofphosphatidylserine and 0.005 mg/ml diacylglycerol, was purchased fromMillipore. Reaction buffer was consisted of 20 mM Hepes pH7.4, 5 mMMgCl₂, and 0.03% Triton X-100. After 2-3 hours reaction time, the assayreadout was developed with KinaseGlo Plus reagent from Promega.Representative compounds of the foregoing examples were tested by thePKCE assay and found to exhibit an IC₅₀ as shown below, where (+)represents an IC₅₀ greater than or equal to 25 μM, (++) represents anIC₅₀ greater than or equal to 10 μM but less than 25 μM, (+++)represents an IC₅₀ greater than or equal to 1 μM but less than 10 μM,and (++++) represents an IC₅₀ less than 1 μM.

Example PKCε IC50 (μM) Flt3 IC50 (μM) KDR IC50 (μM) 118 ++++ ++++ 119++++ +++ 120 +++ 121 + ++++ 122 ++++ 123 ++++ +++ 124 ++ ++ 125 ++ +++126 ++++ +++ 127 +++ + 128 + 129 +++ 130 ++++ +++ 131 + ++++ 132 ++++++++ 133 + +++ 134 +++ +++ 135 ++++ +++ 136 +++ 137 ++++ 138 + 139 ++++140 +++ 141 +++ 142 +++ 143 + 144 +++ 145 + 146 ++++ 147 ++++ 148 ++ 149+++ 150 +++ 151 + 152 ++++ 153 + 154 ++++ 155 +++ 156 +++ 157 + 158 +159 + ++++ 160 + 161 + ++++ 163 ++ 164 ++ 165 ++ 166 ++ 171 ++ 173 ++++++ 174 +++ + 175 +++ + 175 +++ + 176 ++++ +++ 177 ++++ 178 ++++ 179 ++++180 ++++ 182 +++ +++ 183 ++ +++ 184 ++++ ++ 185 +++ +++ 186 +++ +++ 187++++ + 188 ++++ +++ 189 ++++ 190 + 191 ++++ 192 + 193 ++++ 194 + 195++++ + 196 +++ + 199 +++ +++ 201 ++++ ++ 202 ++++ + 203 ++++ + 204 ++++204 ++++ +++ 205 ++++ +++ 206 ++++ +++ 211 ++++ +++ 212 ++++ ++ 213+++ + 214 ++++ ++ 215 +++ + 216 +++ + 217 ++ + 217 ++++ ++ 218 +++ + 219+++ + 222 ++++ + 223 ++++ + 223 ++++ + 224 +++ + 225 +++ + 226 +++ + 227+++ + 228 ++++ ++ 229 +++ + 230 ++++ + 231 ++ ++ 232 ++ ++ 233 +++ +++235 ++++ ++ 236 + ++ 237 +++ ++++ 238 + ++++ 239 ++++ 240 ++++ 241 +++242 + 243 +++ + 244 +++ ++ 249 ++ 261 +++ 264 ++++ 265 ++ 268 +++ 275 ++304 +++ ++++ 312 ++ 320 ++ + 322 +++ + 332 +++ + 333 ++ + 336 ++ + 340+++ ++ 341 ++++ +++ 342 +++ +++ 343 +++ +++ 344 ++ + 345 ++++ ++++ 346++++ 347 ++++ 348 ++++ 349 +++ 350 ++++ 351 ++++ 352 ++++ 353 ++++ 354+++ 355 +++ 356 +++ 357 +++ 359 ++++ ++++ 360 +++ ++++ 361 ++++ ++++ 362++++ ++++ 363 ++++ ++++ 364 ++++ ++++ 364 ++++ +++ 365 ++++ ++++ 366++++ ++++ 367 ++++ ++++ 368 ++++ ++++ 369 ++++ ++++ 370 ++++ ++++ 371+++ 372 ++++ ++++ 373 ++++ ++++ 374 ++++ +++ 375 ++++ +++ 376 ++++ + 377++++ +++ 378 ++++ + 379 ++++ +++ 380 ++++ ++++ 381 ++++ +++ 382 ++++ +++383 ++++ +++ 384 ++++ +++ 385 ++++ +++ 386 ++++ +++ 387 +++ ++ 388 ++++++++ 389 ++++ +++ 390 ++++ +++ 391 +++ + 400 +++ ++++ 400 +++ 401 ++++402 +++ 403 ++++ 404 +++ 405 ++ 406 +++ + 410 +++ + 411 ++ + 418 ++ +422 ++ + 424 +++ + 425 +++ ++ 427 ++ 431 +++ + 433 +++ ++ 434 ++ + 435++ ++++ 436 +++ +++ 437 ++++ +++ 438 + +++ 439 +++ +++ 440 + +++ 441 ++++++ 447 ++ 454 ++ 455 ++ 482 + ++ 483 ++ + 484 ++ + 485 + ++ 487 ++ 505++ 519 ++ 520 +++ 521 +++ 522 +++ 523 ++ 524 +++ 534 +++ 535 ++ 538 ++542 ++ 543 ++ 544 ++ 545 ++ 547 ++ 549 ++ 550 ++ 552 +++ +++ 590 +++++++ 592 +++ 593 + 594 + 595 + 596 + 597 +++ + 598 +++ +++ 612 +++ 614+++ 629 ++ 644 ++ 648 ++ 649 ++ 650 ++

Example 763 IC₅₀ and EC₅₀ Activity of Compounds of the Invention

Using the procedures of Examples 756 (Pim1 ATP depletion assay), 757(Pim2 ATP depletion assay), and 758 (Pim3 ATP depletion assay), the IC₅₀concentration of compounds of the previous examples were determined asshown in the following table, where (+) represents an IC₅₀ greater thanor equal to 25 μM, (++) represents an IC₅₀ greater than or equal to 10μM but less than 25 μM, (+++) represents an IC₅₀ greater than or equalto 1 μM but less than 10 μM, and (++++) represents an IC₅₀ less than 1μM.

Using the procedures of Example 761 (cell proliferation assay), the EC₅₀concentration of compounds of the previous examples in were determinedin HEL 92.1.7, MV4-11 cells and PC3 cells as shown in the followingtable, where (+) represents an EC₅₀ greater than 10 μM, (++) representsan EC₅₀ greater than 5 μM but less than or equal to 10 μM, (+++)represents an EC₅₀ greater than 1 μM but less than or equal to 5 μM, and(++++) represents an EC₅₀ less than or equal to 1 μM.

IC50 (μM) EC50 (μM) Ex. HEL No PIM1 PIM2 PIM3 92.1.7 MV-4-11 PC3 KMS11MM1.s 118 ++++ +++ ++++ 119 ++++ ++++ ++++ 120 ++++ +++ ++++ +++ 121++++ ++++ ++++ 122 ++++ +++ +++ ++++ 123 ++++ +++ ++++ ++++ 124 +++ ++++ 125 +++ + ++ 126 ++++ +++ ++++ ++ 127 +++ + + 128 +++ + ++ 130 ++++++++ ++++ ++ 131 ++++ ++++ ++++ ++++ 132 ++++ ++++ ++++ 133 ++++ ++++++++ 134 ++++ ++++ ++++ 135 ++++ ++++ ++++ 136 ++++ ++++ ++++ 137 ++++++++ ++++ 138 ++++ ++++ ++++ 139 ++++ ++++ ++++ 140 ++++ ++++ ++++ 141++++ ++++ ++++ 142 ++++ ++++ ++++ +++ 143 ++++ ++++ ++++ +++ 145 ++++++++ ++++ +++ 146 ++++ ++++ ++++ + 147 ++++ ++++ ++++ ++ 148 ++++ ++++++++ + 150 ++++ ++++ ++++ +++ 151 ++++ ++++ ++++ + 152 ++++ ++++ ++++153 ++++ ++++ ++++ 154 ++++ ++++ ++++ 155 ++++ ++++ ++++ ++ 156 ++++++++ ++++ 157 ++++ ++++ ++++ + 158 ++++ ++++ ++++ 159 ++++ ++++ ++++ ++160 ++++ ++++ ++++ + 161 ++++ ++++ ++++ + 162 ++++ ++++ ++++ 163 ++++++++ ++++ 164 ++++ ++++ ++++ 165 ++++ ++++ ++++ 166 ++++ ++++ ++++ 167++++ +++ ++++ 168 ++++ ++++ ++++ 169 ++++ ++++ ++++ 170 ++++ +++ ++++171 ++++ +++ ++++ 173 ++++ ++++ ++++ +++ ++ 174 ++++ ++++ ++++ + 177++++ ++++ ++++ + 175 ++++ ++++ ++++ + 176 ++++ ++++ ++++ +++ +++ 177++++ ++++ ++++ 178 ++++ ++++ ++++ 179 ++++ ++++ ++++ +++ 180 ++++ ++++++++ ++++ 181 ++++ ++++ ++++ ++++ 182 ++++ ++++ ++++ ++++ 183 ++++ ++++++++ ++++ 184 ++++ +++ ++++ ++++ 185 ++++ ++++ ++++ ++++ 186 ++++ ++++++++ 187 ++++ ++++ ++++ +++ ++++ +++ 188 ++++ +++ ++++ 189 ++++ +++ ++++191 ++++ + +++ 192 + + + 193 ++++ +++ ++++ ++++ 194 + + + 195 ++++ ++++++++ + 196 +++ ++ +++ 197 + + + 198 + + + 199 +++ + + 199 +++ + ++ 199+++ + ++ 200 ++ + + 201 ++++ ++++ ++++ ++ 202 +++ + +++ +++ 203 ++++ +++++++ ++++ 204 ++++ ++++ ++++ 205 ++++ ++++ ++++ ++++ 206 ++++ +++ ++++207 ++++ ++ ++++ ++ 208 ++++ +++ ++++ ++++ 209 ++ + + +++ 210 ++++ +++++++ ++++ 211 ++++ ++++ ++++ 212 ++++ ++++ ++++ 213 +++ + +++ 214 +++++++ ++++ +++ 215 ++++ +++ ++++ 216 +++ + +++ 217 ++++ + +++ ++ 218 ++++++ ++++ 219 ++++ ++ ++++ 220 ++++ ++ ++++ 221 ++++ +++ ++++ 222 ++++ ++++++ + 223 ++++ +++ ++++ 224 ++++ + +++ 225 ++++ ++ ++++ 226 ++++ +++++++ 227 +++ + +++ 228 ++++ ++ ++++ ++ 229 ++++ +++ ++++ 230 ++++ ++++++++ 231 +++ + +++ 232 ++ + ++ 233 +++ + +++ 234 +++ + ++ 235 ++++ ++++++++ + 236 ++++ ++++ ++++ 237 ++++ ++++ ++++ 238 ++++ ++++ ++++ 239 ++++++++ ++++ + 240 ++++ ++++ ++++ 241 ++++ ++++ ++++ 242 ++++ ++++ ++++ 243+++ ++ +++ 244 +++ ++ +++ 245 + + ++ 246 ++++ ++++ ++++ 247 ++++ ++++++++ 248 ++++ ++++ ++++ 249 ++++ ++++ ++++ 250 ++++ ++++ ++++ 251 ++++++++ ++++ 252 ++++ ++++ ++++ 253 ++++ ++++ ++++ 254 ++++ ++++ ++++ 255++++ ++++ ++++ 256 ++++ ++++ ++++ 257 ++++ ++++ ++++ 258 ++++ ++++ ++++259 ++++ ++++ ++++ 260 ++++ ++++ ++++ 261 ++++ ++++ ++++ 262 ++++ +++++++ 263 ++++ ++++ ++++ 264 ++++ ++++ ++++ 265 ++++ ++++ ++++ 266 ++++++++ ++++ 267 ++++ ++++ ++++ 268 ++++ ++++ ++++ 269 ++++ ++++ ++++ 270++++ ++++ ++++ 271 ++++ ++++ ++++ 272 ++++ ++++ ++++ 273 ++++ ++++ ++++274 ++++ ++++ ++++ 275 ++++ ++++ ++++ 276 ++++ ++++ ++++ +++ ++ 277 ++++++++ ++++ 278 ++++ ++++ ++++ 279 ++++ ++++ ++++ 280 ++++ ++++ ++++ 281++++ ++++ ++++ 282 ++++ ++++ ++++ 283 ++++ ++++ ++++ 284 ++++ ++++ ++++285 ++++ ++++ ++++ 286 ++++ ++++ ++++ 287 ++++ ++++ ++++ 288 ++++ ++++++++ 289 ++++ ++++ ++++ 290 ++++ ++++ ++++ +++ +++ 291 ++++ ++++ ++++292 ++++ ++++ ++++ ++++ ++++ 293 ++++ ++++ ++++ 294 ++++ ++++ ++++ 295++++ +++ ++++ 296 ++++ ++++ ++++ 297 ++++ ++++ ++++ 298 ++++ ++++ ++++++++ ++++ 299 ++++ ++++ ++++ 300 ++++ ++++ ++++ ++++ ++++ 301 ++++ ++++++++ 302 ++++ ++++ ++++ 303 ++++ ++++ ++++ 304 ++++ + +++ 312 ++++ +++++++ 317 ++++ ++++ ++++ +++ + +++ ++++ 318 ++++ ++++ ++++ 319 ++++ ++++++++ +++ 320 ++++ ++++ ++++ + 321 ++++ ++++ ++++ +++ 322 ++++ ++++ ++++++++ ++ 323 ++++ ++++ ++++ ++++ +++ 324 ++++ ++++ ++++ ++++ 325 ++++++++ ++++ +++ 326 ++++ ++++ ++++ +++ 327 ++++ ++++ ++++ +++ 328 ++++++++ ++++ +++ ++++ ++++ 329 ++++ ++++ ++++ ++ 330 ++++ ++++ ++++ + 331++++ ++++ ++++ +++ 332 ++++ ++++ ++++ +++ 333 ++++ ++++ ++++ +++ 334++++ ++++ ++++ ++ 335 ++++ ++++ ++++ ++ 336 ++++ ++++ ++++ 337 ++++ ++++++++ ++ 338 ++++ ++++ ++++ 339 ++++ ++++ ++++ +++ 340 ++++ ++++ ++++ +++341 ++++ ++++ ++++ +++ 342 ++++ ++++ ++++ ++++ 343 ++++ ++++ ++++ ++++344 ++++ ++++ ++++ + 345 ++++ ++++ ++++ ++++ ++++ 346 ++++ ++++ ++++++++ 347 ++++ ++++ ++++ ++++ 348 ++++ ++++ ++++ ++++ +++ 349 ++++ ++++++++ ++++ 350 ++++ ++++ ++++ ++++ 351 ++++ ++++ ++++ 352 ++++ ++++ ++++++++ 353 ++++ ++++ ++++ ++++ 355 ++++ ++++ ++++ + 356 ++++ ++++ ++++ +357 ++++ ++++ ++++ +++ 358 ++++ ++++ ++++ ++++ 359 ++++ ++++ ++++ ++++360 +++ + +++ 361 ++++ ++++ ++++ 362 ++++ ++++ ++++ ++++ 363 ++++ ++++++++ ++++ 364 ++++ ++++ ++++ ++++ 365 ++++ +++ ++++ + 366 ++++ +++ ++++367 ++++ ++++ ++++ ++++ 368 ++++ ++++ ++++ ++++ 369 ++++ ++++ ++++ 370++++ ++++ ++++ ++ 371 ++++ ++++ ++++ 372 ++++ ++++ ++++ +++ 376 ++++++++ ++++ + 377 ++++ ++++ ++++ 378 ++++ ++ +++ +++ 379 ++++ +++ ++++ 380++++ +++ ++++ 381 ++++ ++++ ++++ 382 ++++ +++ ++++ +++ 383 ++++ +++ ++++384 ++++ +++ +++ 385 ++++ +++ ++++ 386 ++++ +++ ++++ 387 +++ + +++ 388++++ +++ ++++ 389 ++++ +++ +++ +++ 390 ++++ + +++ 391 +++ + +++ 393 ++++++++ ++++ +++ 395 ++++ + +++ +++ 396 ++++ +++ ++++ +++ 397 ++++ ++++++++ ++++ 398 +++ ++ +++ 399 ++++ ++++ ++++ ++++ 401 ++++ ++++ ++++ ++402 ++++ ++++ ++++ ++ 403 ++++ ++++ ++++ 404 ++++ ++++ ++++ 405 ++++++++ ++++ 406 ++++ ++++ ++++ 407 ++++ ++++ ++++ 408 ++++ ++++ ++++ 409++++ ++++ ++++ 410 ++++ ++++ ++++ +++ 411 ++++ ++++ ++++ 412 ++++ ++++++++ ++ 413 ++++ ++++ ++++ ++++ 414 ++++ ++++ ++++ +++ 415 ++++ ++++++++ ++++ 416 ++++ ++++ ++++ 417 ++++ ++++ ++++ 418 ++++ ++++ ++++ +++419 ++++ ++++ ++++ +++ 420 ++++ ++++ ++++ 421 ++++ ++++ ++++ +++ + 422++++ ++++ ++++ +++ + 423 ++++ ++++ ++++ +++ 424 ++++ ++++ ++++ +++ ++425 ++++ ++++ ++++ ++++ ++ ++++ ++++ 426 ++++ ++++ ++++ ++ 427 ++++ ++++++++ +++ 428 ++++ ++++ ++++ ++ 429 ++++ ++++ ++++ ++ 430 ++++ ++++ +++++++ 431 ++++ ++++ ++++ +++ 432 ++++ ++++ ++++ +++ 433 ++++ ++++ ++++ +++++++ 434 ++++ ++++ ++++ +++ ++++ 435 +++ +++ +++ 436 +++ + +++ 437 ++++++++ ++++ 438 +++ + +++ 439 +++ + +++ 440 + + ++ 441 +++ + +++ 442 ++++++++ ++++ +++ ++ 443 ++++ ++++ ++++ +++ ++ 444 ++++ ++++ ++++ 445 ++++++++ ++++ +++ +++ 446 ++++ ++++ ++++ 447 ++++ ++++ ++++ 448 ++++ ++++++++ 449 ++++ ++++ ++++ 450 ++++ ++++ ++++ 451 ++++ ++++ ++++ 452 ++++++++ ++++ 453 ++++ ++++ ++++ 454 ++++ ++++ ++++ +++ ++ 455 ++++ ++++++++ 456 ++++ ++++ ++++ 457 ++++ ++++ ++++ 458 ++++ ++++ ++++ 459 ++++++++ ++++ 460 ++++ ++++ ++++ 461 ++++ ++++ ++++ 462 ++++ ++++ ++++ ++++++ 463 ++++ ++++ ++++ 464 ++++ ++++ ++++ 465 ++++ ++++ ++++ 466 ++++++++ ++++ 467 ++++ ++++ ++++ 468 ++++ ++++ ++++ 469 ++++ ++++ ++++ 470++++ ++++ ++++ 471 ++++ ++++ ++++ 472 ++++ ++++ ++++ 473 ++++ ++++ ++++474 ++++ ++++ ++++ 475 ++++ ++++ ++++ 476 ++++ ++++ ++++ 477 ++++ ++++++++ 478 ++++ ++++ ++++ 479 ++++ ++++ ++++ 480 ++++ ++++ ++++ 481 ++++++++ ++++ 482 ++++ ++++ ++++ 483 ++++ ++++ ++++ 484 ++++ ++++ ++++ 485++++ ++++ ++++ 486 ++++ ++++ ++++ 487 ++++ ++++ ++++ 488 ++++ ++++ ++++489 ++++ ++++ ++++ 490 ++++ ++++ ++++ 491 ++++ ++++ ++++ +++ +++ 492++++ ++++ ++++ 493 ++++ ++++ ++++ 494 ++++ ++++ ++++ ++++ +++ 495 ++++++++ ++++ 496 ++++ ++++ ++++ ++++ ++++ 497 ++++ ++++ ++++ 498 ++++ ++++++++ 499 ++++ ++++ ++++ 500 ++++ ++++ ++++ 501 ++++ ++++ ++++ 502 ++++++++ ++++ 503 ++++ ++++ ++++ 504 ++++ ++++ ++++ 505 ++++ ++++ ++++ 506++++ ++++ ++++ 507 ++++ ++++ ++++ 508 ++++ ++++ ++++ 509 ++++ ++++ ++++510 ++++ ++++ ++++ 511 ++++ ++++ ++++ 512 ++++ ++++ ++++ 513 ++++ ++++++++ 514 ++++ ++++ ++++ 515 ++++ ++++ ++++ 516 ++++ ++++ ++++ ++++ ++++517 ++++ ++++ ++++ +++ +++ 519 ++++ ++++ ++++ 520 ++++ ++++ ++++ ++ ++521 ++++ ++++ ++++ 522 ++++ ++++ ++++ 523 ++++ ++++ ++++ 524 ++++ ++++++++ 525 ++++ ++++ ++++ 526 ++++ ++++ ++++ 527 ++++ ++++ ++++ 528 ++++++++ ++++ 529 ++++ ++++ ++++ 530 ++++ ++++ ++++ 531 ++++ ++++ ++++ 532++++ ++++ ++++ 533 ++++ ++++ ++++ 534 ++++ ++++ ++++ 535 ++++ ++++ ++++536 ++++ ++++ ++++ 537 ++++ ++++ ++++ 538 ++++ ++++ ++++ 539 ++++ ++++++++ 540 ++++ ++++ ++++ 541 ++++ ++++ ++++ 542 ++++ ++++ ++++ 543 ++++++++ ++++ 544 ++++ ++++ ++++ 545 ++++ ++++ ++++ 546 ++++ ++++ ++++ 547++++ ++++ ++++ 548 ++++ ++++ ++++ 549 ++++ ++++ ++++ 550 ++++ ++++ ++++551 ++++ ++++ ++++ 552 ++++ ++++ ++++ 553 ++++ ++++ ++++ 554 ++++ ++++++++ 555 ++++ ++++ ++++ 556 ++++ ++++ ++++ 557 ++++ ++++ ++++ 558 ++++++++ ++++ 559 ++++ ++++ ++++ 560 ++++ ++++ ++++ 561 ++++ ++++ ++++ 562++++ ++++ ++++ 563 ++++ ++++ ++++ 564 ++++ ++++ ++++ 565 ++++ ++++ ++++566 ++++ ++++ ++++ 567 ++++ ++++ ++++ 568 ++++ ++++ ++++ 569 ++++ ++++++++ 570 ++++ ++++ ++++ +++ +++ 571 ++++ ++++ ++++ 572 ++++ ++++ ++++573 ++++ ++++ ++++ 574 ++++ ++++ ++++ 575 ++++ ++++ +++ 576 ++++ ++++++++ 577 ++++ ++++ ++++ +++ +++ 578 ++++ ++++ ++++ 579 ++++ ++++ ++++++++ +++ 580 ++++ ++++ ++++ +++ +++ 581 ++++ ++++ ++++ 582 ++++ +++ ++++584 ++++ +++ ++++ 585 ++++ ++++ ++++ 586 ++++ +++ ++++ 587 ++++ ++++++++ ++++ 588 ++++ ++++ ++++ 589 ++++ ++++ ++++ 590 ++++ ++++ ++++ +++591 ++++ ++++ ++++ ++ 592 ++++ ++++ ++++ ++++ 593 ++++ ++++ ++++ 594++++ ++++ ++++ 595 ++++ ++++ ++++ 596 ++++ ++++ ++++ 597 ++++ +++ ++++598 ++++ +++ ++++ 599 ++++ +++ +++ 600 ++++ +++ ++++ 601 ++++ ++++ ++++602 ++++ +++ ++++ 603 ++++ ++++ ++++ 604 ++++ ++++ ++++ 605 ++++ ++++++++ 606 ++++ ++++ ++++ 607 ++++ ++++ ++++ 608 ++++ ++++ ++++ 609 ++++++++ ++++ 610 ++++ ++++ ++++ 611 ++++ ++++ ++++ 612 ++++ +++ ++++ 613++++ ++++ ++++ 614 ++++ ++++ ++++ 615 ++++ ++++ ++++ 616 ++++ ++++ ++++617 ++++ ++++ ++++ 618 ++++ ++++ ++++ 619 ++++ ++++ ++++ 620 ++++ ++++++++ 621 ++++ ++++ ++++ 622 ++++ ++++ ++++ 623 ++++ ++++ ++++ 624 ++++++++ ++++ 625 ++++ ++++ ++++ 626 ++++ ++++ ++++ 627 ++++ ++++ ++++ 628++++ ++++ ++++ 629 ++++ ++++ ++++ 630 ++++ ++++ ++++ 631 ++++ ++++ ++++632 ++++ ++++ ++++ 633 ++++ ++++ ++++ 634 ++++ ++++ ++++ 635 ++++ ++++++++ 636 ++++ ++++ ++++ 637 ++++ ++++ ++++ 638 ++++ ++++ ++++ 639 ++++++++ ++++ 640 ++++ ++++ ++++ 641 ++++ ++++ ++++ 642 ++++ ++++ ++++ 643++++ ++++ ++++ 644 ++++ ++++ ++++ 645 ++++ ++++ ++++ 646 ++++ ++++ ++++647 ++++ ++++ ++++ 648 ++++ ++++ ++++ 649 ++++ ++++ ++++ 650 ++++ ++++++++ 651 ++++ ++++ ++++ 652 ++++ ++++ ++++ 653 ++++ ++++ ++++ 654 ++++++++ ++++ 655 ++++ ++++ ++++ 656 ++++ ++++ ++++ 657 ++++ ++++ ++++ 658++++ ++++ ++++ 659 ++++ ++++ ++++ 660 ++++ ++++ ++++ 661 ++++ ++++ ++++662 ++++ ++++ ++++ 663 ++++ ++++ ++++ 664 ++++ ++++ ++++ 665 ++++ ++++++++ 666 ++++ ++++ ++++ 667 ++++ ++++ ++++ 668 ++++ ++++ ++++ 669 ++++++++ ++++ 670 ++++ ++++ ++++ 671 ++++ ++++ ++++ 672 ++++ ++++ ++++ 673++++ ++++ ++++ 674 ++++ ++++ ++++ 675 ++++ ++++ ++++ 676 ++++ ++++ ++++677 ++++ ++++ ++++ 678 ++++ ++++ ++++ 679 ++++ ++++ ++++ +++ ++++ 680++++ ++++ ++++ 681 ++++ ++++ ++++ 682 ++++ ++++ ++++ 683 ++++ ++++ ++++++++ +++ 684 ++++ ++++ ++++ 685 ++++ ++++ ++++ 686 ++++ ++++ ++++ 687++++ ++++ ++++ 688 ++++ ++++ ++++ 689 ++++ ++++ ++++ 690 ++++ ++++ ++++691 ++++ ++++ ++++ 692 ++++ ++++ ++++ 693 ++++ ++++ ++++ 694 ++++ ++++++++ 695 ++++ ++++ ++++ 696 ++++ ++++ ++++ 697 ++++ +++ ++++ 698 ++++++++ ++++ 699 ++++ ++++ ++++ 700 ++++ ++++ ++++ 701 ++++ ++++ ++++ 702++++ ++++ ++++ +++ +++ 703 ++++ ++++ ++++ 704 ++++ ++++ ++++ 705 ++++++++ ++++ 706 ++++ ++++ ++++ 707 ++++ ++++ ++++ 708 ++++ ++++ ++++ 709++++ ++++ ++++ 710 ++++ ++++ ++++ 711 ++++ ++++ ++++ 712 ++++ ++++ ++++713 ++++ ++++ ++++ 714 ++++ ++++ ++++ 715 ++++ ++++ ++++ 716 ++++ ++++++++ 717 ++++ ++++ ++++ 718 ++++ ++++ ++++ 719 ++++ ++++ ++++ 720 ++++++++ ++++ 721 ++++ ++++ ++++ 722 ++++ ++++ ++++ 723 ++++ ++++ ++++ 724++++ ++++ ++++ 725 ++++ ++++ ++++ 726 ++++ ++++ ++++ 727 ++++ ++++ ++++728 ++++ ++++ ++++ 729 ++++ ++++ ++++ 730 ++++ ++++ ++++ 731 ++++ ++++++++ 732 ++++ ++++ ++++ 733 ++++ ++++ ++++ 734 ++++ ++++ ++++ 735 ++++++++ ++++ 736 ++++ ++++ ++++ ++++ +++ 737 ++++ ++++ ++++ 738 ++++ ++++++++ 739 ++++ ++++ ++++ 740 ++++ ++++ ++++ 741 ++++ ++++ ++++ 742 ++++++++ ++++ 743 ++++ ++++ ++++ 744 ++++ ++++ ++++ +++ +++ 745 ++++ ++++++++ 746 ++++ ++++ ++++ 747 ++++ ++++ ++++ 748 ++++ ++++ ++++ 749 ++++++++ ++++ 750 ++++ ++++ ++++ 751 ++++ ++++ ++++ 752 ++++ ++++ ++++ 753++++ ++++ ++++ +++ ++++ 754 ++++ ++++ ++++ ++++ +++ 755 ++++ ++++ ++++

While illustrative embodiments have been illustrated and described, itwill be appreciated that various changes can be made therein withoutdeparting from the spirit and scope of the invention.

1. A compound of Formula I, or a stereoisomer, tautomer, orpharmaceutically acceptable salt thereof,

wherein, X₁, X₂, X₃ and X₄ are CR₂ or X₃ and X₁ are N and X₂ is CR₂; Yis substituted or unsubstituted amino, alkoxy, cycloalkyl,heterocycloalkyl, aryl or heteroaryl; Z₁, Z₂ and Z₃ are independentlyselected from CR₂ and N; provided that not more than one of Z₁, Z₂ andZ₃ can be N; R₁ is selected from the group consisting of hydrogen, halo,alkyl, cycloalkyl, —CN, —NO₂, and —NHR₃; each R₂ is independentlyselected from the group consisting of hydrogen, halo, hydroxyl, nitro,cyano, SO₃H and substituted or unsubstituted alkyl, alkenyl, alkynyl,alkoxy, amino, aminocarbonyl, aminothiocarbonyl, aminocarbonylamino,aminothiocarbonylamino, aminocarbonyloxy, aminosulfonyl,aminosulfonyloxy, aminosulfonylamino, amidino, carboxyl, carboxyl ester,(carboxyl ester)amino, (carboxyl ester)oxy, sulfonyl, sulfonyloxy,thioacyl, thiol, alkylthio, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, partially saturated cycloalkyl, aryloxy, heteroaryloxy,heterocyclyloxy, cycloalkyloxy, acyl, acylamino and acyloxy; R₃ isselected from the group consisting of hydrogen, —CO—R₄ and substitutedor unsubstituted alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;and R₄ is selected from the group consisting of alkyl, substitutedalkyl, alkoxy, substituted alkoxy, amino, substituted amino, andalkylamino.
 2. A compound of claim 1 wherein X₁, X₂, X₃ and X₄ are CR₂and Z₂ is N and Z₁ and Z₃ are CR₂. A compound of claim 1 wherein X₁ is Nand X₂, X₃ and X₄ are CR₂.
 3. A compound of claim 1 wherein X₁, X₂, X₃and X₄ are CR₂ and Z₃ is N and Z₁ and Z₂ are CR₂.
 4. A compound of claim1 wherein Y is substituted or unsubstituted piperidinyl or piperazinyl.5. A compound of claim 1 wherein Z₁, Z₂ and Z₃ are CR₂.
 6. A compound ofFormula I, or a stereoisomer, tautomer, or pharmaceutically acceptablesalt thereof,

wherein, X₄ is N and X₁, X₂ and X₃ are CR₂ or X₃ is N and X₁, X₂ and X₄are CR₂; Y is substituted or unsubstituted amino, alkoxy, cycloalkyl,heterocycloalkyl, aryl or heteroaryl; Z₁, Z₂ and Z₃ are independentlyselected from CR₂ and N; provided that not more than one of Z₁, Z₂ andZ₃ can be N; R₁ is selected from the group consisting of hydrogen, halo,alkyl, cycloalkyl, —CN, —NO₂, and —NHR₃; each R₂ is independentlyselected from the group consisting of hydrogen, halo, hydroxyl, nitro,cyano, SO₃H and substituted or unsubstituted alkyl, alkenyl, alkynyl,alkoxy, amino, aminocarbonyl, aminothiocarbonyl, aminocarbonylamino,aminothiocarbonylamino, aminocarbonyloxy, aminosulfonyl,aminosulfonyloxy, aminosulfonylamino, amidino, carboxyl, carboxyl ester,(carboxyl ester)amino, (carboxyl ester)oxy, sulfonyl, sulfonyloxy,thioacyl, thiol, alkylthio, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, partially saturated cycloalkyl, aryloxy, heteroaryloxy,heterocyclyloxy, cycloalkyloxy, acyl, acylamino and acyloxy; R₃ isselected from the group consisting of hydrogen, —CO—R₄ and substitutedor unsubstituted alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;and R₄ is selected from the group consisting of alkyl, substitutedalkyl, alkoxy, substituted alkoxy, amino, substituted amino, andalkylamino.
 7. A compound of claim 6 wherein Z₁, Z₂ and Z₃ are CR₂.
 8. Acompound of claim 6 wherein Z₁ is N and Z₂ and Z₃ are CR₂.
 9. A compoundof claim 6 wherein Z₂ is N and Z₁ and Z₃ are CR₂.
 10. A compound ofclaim 6 wherein Z₃ is N and Z₁ and Z₂ are CR₂.
 11. A compositioncomprising a therapeutically effective amount of compound of claim 1, ora stereoisomer, tautomer, or pharmaceutically acceptable salt thereof,together with a pharmaceutically acceptable carrier.
 12. A method fortreating a cancer disorder in a patient, comprising administering to thepatient a composition comprising an amount of a compound of claim
 1. 13.The composition of claim 11 which further comprises at least oneadditional agent for the treatment of cancer.
 14. A compositioncomprising a therapeutically effective amount of compound of claim 6, ora stereoisomer, tautomer, or pharmaceutically acceptable salt thereof,together with a pharmaceutically acceptable carrier.
 15. A method fortreating a cancer disorder in a patient, comprising administering to thepatient a composition comprising an amount of a compound of claim
 6. 16.The composition of claim 14 which further comprises at least oneadditional agent for the treatment of cancer.